36 results on '"Takatsugu Kan"'
Search Results
2. Safety assessment of robotic gastrectomy and analysis of surgical learning process: a multicenter cohort study
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Norihiro, Shimoike, Tatsuto, Nishigori, Yoshito, Yamashita, Masato, Kondo, Dai, Manaka, Yoshio, Kadokawa, Atsushi, Itami, Seiichiro, Kanaya, Hisahiro, Hosogi, Seiji, Satoh, Hiroaki, Hata, Takatsugu, Kan, Hironori, Kawada, Michihiro, Yamamoto, Eiji, Tanaka, Shigeru, Tsunoda, Shigeo, Hisamori, Koya, Hida, Kentaro, Ueno, Shiro, Tanaka, and Kazutaka, Obama
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Cohort Studies ,Postoperative Complications ,Treatment Outcome ,Robotic Surgical Procedures ,Gastrectomy ,Stomach Neoplasms ,Humans ,Laparoscopy ,Retrospective Studies - Abstract
The safety of robotic gastrectomy (RG) for gastric cancer in daily clinical settings and the process by which surgeons are introduced and taught RG remain unclear. This study aimed to evaluate the safety of RG in daily clinical practice and assess the learning process in surgeons introduced to RG.Patients who underwent RG for gastric cancer at Kyoto University and 12 affiliated hospitals across Japan from January 2017 to October 2019 were included. Any morbidity with a Clavien-Dindo classification grade of II or higher was evaluated. Moreover, the influence of the surgeon's accumulated RG experience on surgical outcomes and surgeon-reported postoperative fatigue were assessed.A total of 336 patients were included in this study. No conversion to open or laparoscopic surgery and no in-hospital mortality were observed. Overall, 50 (14.9%) patients developed morbidity. During the study period, 14 surgeons were introduced to robotic procedures. The initial five cases had surprisingly lower incidence of morbidity compared to the following cases (odds ratio 0.29), although their operative time was longer (+ 74.2 min) and surgeon's fatigue scores were higher (+ 18.4 out of 100 in visual analog scale).RG was safely performed in actual clinical settings. Although the initial case series had longer operative time and promoted greater levels of surgeon fatigue compared to subsequent cases, our results suggested that RG had been introduced safely.
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- 2021
3. Medial Approach for Laparoscopic Total Gastrectomy with Splenic Lymph Node Dissection
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Takatsugu Kan, Atsushi Itami, Kazutaka Obama, Yoshiharu Sakai, Hiroshi Okabe, and Eiji Tanaka
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Adult ,Male ,medicine.medical_specialty ,Treatment outcome ,MEDLINE ,Dissection (medical) ,Gastrectomy ,Stomach Neoplasms ,Medial approach ,medicine ,Humans ,Laparoscopic total gastrectomy ,Aged ,Aged, 80 and over ,business.industry ,Middle Aged ,medicine.disease ,Surgery ,Treatment Outcome ,medicine.anatomical_structure ,Lymph Node Excision ,Female ,Laparoscopy ,business ,Spleen ,Splenic lymph nodes - Published
- 2010
4. Chromosomal abnormalities and novel disease-related regions in progression from Barrett's esophagus to esophageal adenocarcinoma
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Harmik J. Soukiasian, Motohiro Kato, Tadayuki Akagi, Seishi Ogawa, Tetsuo Ito, Go Yamamoto, Carl W. Miller, Jessica K.R. Boult, Stephen J. Meltzer, H. Phillip Koeffler, Yulan Cheng, Takatsugu Kan, Alexandru Olaru, Zhe Jin, and Norihiko Kawamata
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Male ,Cancer Research ,Pathology ,medicine.medical_specialty ,Esophageal Neoplasms ,Tumor suppressor gene ,Blotting, Western ,Gene Dosage ,Loss of Heterozygosity ,Adenocarcinoma ,Biology ,Polymorphism, Single Nucleotide ,Article ,Loss of heterozygosity ,Barrett Esophagus ,Esophagus ,CDKN2A ,Biomarkers, Tumor ,medicine ,Humans ,RNA, Messenger ,Aged ,Oligonucleotide Array Sequence Analysis ,Aged, 80 and over ,Chromosome Aberrations ,Genome, Human ,Reverse Transcriptase Polymerase Chain Reaction ,Esophageal disease ,Gene Expression Profiling ,Cytogenetics ,Middle Aged ,medicine.disease ,Molecular biology ,DNA-Binding Proteins ,medicine.anatomical_structure ,Oncology ,Tumor progression ,Barrett's esophagus ,Disease Progression ,Female ,Genome-Wide Association Study - Abstract
Barrett’s esophagus (BE) is a metaplastic condition caused by chronic gastroesophageal reflux which represents an early step in the development of esophageal adenocarcinoma (EAC). Single-nucleotide polymorphism microarray (SNP-chip) analysis is a novel, precise, high-throughput approach to examining genomic alterations in neoplasia. Using 250K SNP-chips, we examined the neoplastic progression of BE to EAC, studying 11 matched sample sets: 6 sets of normal esophagus [NE], BE and EAC, 4 of NE and BE, and 1 of NE and EAC. Six (60%) of 10 total BE samples and 4 (57%) of 7 total EAC samples exhibited one or more genomic abnormalities comprising deletions, duplications, amplifications, and copy-number-neutral loss of heterozygosity (CNN-LOH). Several shared abnormalities were identified, including chromosome 9p CNN-LOH (2 BE samples [20%]), deletion of CDKN2A (4 BE samples [40%]), and amplification of 17q12–21.2 involving the ERBB2, RARA and TOP2A genes (3.1Mb, 2 EAC [29%]). Interestingly, one BE sample contained a homozygous deletion spanning 9p22.3-p22.2 (1.2 Mb): this region harbors only one known gene, basonuclin 2 (BNC2). Real-time PCR analysis confirmed deletion of this gene and decreased expression of BNC2 mRNA in the BE sample. Furthermore, transfection and stable expression of BNC2 caused growth arrest of OE33 EAC cells, suggesting that BNC2 functions as a tumor suppressor gene in the esophagus, and that deletion of this gene occurs during the development of EAC. Thus, this SNP-chip analysis has identified several early cytogenetic events and novel candidate cancer-related genes that are potentially involved in the evolution of BE to EAC.
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- 2009
5. MicroRNA-21 is overexpressed in human cholangiocarcinoma and regulates programmed cell death 4 and tissue inhibitor of metalloproteinase 3
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Michael Torbenson, Alexandru Olaru, Paul J. Thuluvath, Gregory J. Gores, James P. Hamilton, Stephen J. Meltzer, Yulan Cheng, Anirban Maitra, Yuriko Mori, Florin M. Selaru, Rachana Agarwal, Stefan David, Jian Yang, John Abraham, Perumal Vivekanandan, Wayne Yu, Christos S. Georgiades, Bogdan C. Paun, Hector A Alvarez, Ralph H. Hruban, Zhe Jin, Takatsugu Kan, and Nicholas F. LaRusso
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Adult ,Male ,medicine.medical_specialty ,Programmed cell death ,Pathology ,Biology ,Article ,Cholangiocarcinoma ,Cell Line, Tumor ,Internal medicine ,microRNA ,medicine ,Humans ,RNA, Messenger ,Aged ,Oligonucleotide Array Sequence Analysis ,Aged, 80 and over ,Tissue Inhibitor of Metalloproteinase-3 ,Regulation of gene expression ,Hepatology ,Reverse Transcriptase Polymerase Chain Reaction ,Gene Expression Profiling ,RNA-Binding Proteins ,Cancer ,Middle Aged ,Tissue inhibitor of metalloproteinase ,medicine.disease ,Candidate Tumor Suppressor Gene ,Gene Expression Regulation, Neoplastic ,Gene expression profiling ,MicroRNAs ,Bile Ducts, Intrahepatic ,Bile Duct Neoplasms ,Cancer research ,Female ,Apoptosis Regulatory Proteins - Abstract
Cholangiocarcinomas (CCAs) are aggressive cancers, with high mortality and poor survival rates. Only radical surgery offers patients some hope of cure; however, most patients are not surgical candidates because of late diagnosis secondary to relatively poor accuracy of diagnostic means. MicroRNAs (miRs) are involved in every cancer examined, but they have not been evaluated in primary CCA. In this study, miR arrays were performed on five primary CCAs and five normal bile duct specimens (NBDs). Several miRs were dysregulated and miR-21 was overexpressed in CCAs. miR-21 differential expression in these 10 specimens was verified by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). To validate these findings, qRT-PCR for miR-21 was then performed on 18 additional primary CCAs and 12 normal liver specimens. MiR-21 was 95% sensitive and 100% specific in distinguishing between CCA and normal tissues, with an area under the receiver operating characteristic curve of 0.995. Inhibitors of miR-21 increased protein levels of programmed cell death 4 (PDCD4) and tissue inhibitor of metalloproteinases 3 (TIMP3). Notably, messenger RNA levels of TIMP3 were significantly lower in CCAs than in normals. Conclusions: MiR-21 is overexpressed in human CCAs. Furthermore, miR-21 may be oncogenic, at least in part, by inhibiting PDCD4 and TIMP3. Finally, these data suggest that TIMP3 is a candidate tumor suppressor gene in the biliary tree. (HEPATOLOGY 2009.)
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- 2009
6. Aberrant silencing of the endocrine peptide gene tachykinin-1 in gastric cancer
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Stefan David, Yuriko Mori, Yulan Cheng, Takatsugu Kan, Zhe Jin, and Rachana Agarwal
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Adult ,Male ,Biophysics ,Adenocarcinoma ,Biology ,medicine.disease_cause ,Biochemistry ,Stomach Neoplasms ,TAC1 ,Tachykinins ,medicine ,Humans ,Gene silencing ,Gene Silencing ,Protein Precursors ,Promoter Regions, Genetic ,Molecular Biology ,Aged ,Aged, 80 and over ,Regulation of gene expression ,Helicobacter pylori ,Age Factors ,Cancer ,Promoter ,Cell Biology ,Methylation ,DNA Methylation ,Middle Aged ,medicine.disease ,Molecular biology ,Gene Expression Regulation, Neoplastic ,Cell Transformation, Neoplastic ,DNA methylation ,Female ,Microsatellite Instability ,Carcinogenesis - Abstract
Tachykinin-1 (TAC1) is the precursor protein for neuroendocrine peptides, including substance P, and is centrally involved in gastric secretion, motility, mucosal immunity, and cell proliferation. Here we report aberrant silencing of TAC1 in gastric cancer (GC) by promoter hypermethylation. TAC1 methylation and mRNA expression in 47 primary GCs and 41 noncancerous gastric mucosae (NLs) were analyzed by utilizing real-time quantitative PCR-based assays. TAC1 methylation was more prevalent in GCs than in NLs: 21 (45%) of 47 GCs versus 6 (15%) of 41 NLs (p
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- 2009
7. Promoter hypermethylation of CDH13 is a common, early event in human esophageal adenocarcinogenesis and correlates with clinical risk factors
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Fumiaki Sato, Zhe Jin, Rachana Agarwal, Takatsugu Kan, Jian Yang, Tetsuo Ito, John M. Abraham, Alexandru Olaru, Yutaka Shimada, Stephen J. Meltzer, Yuriko Mori, Florin M. Selaru, Stefan David, Yulan Cheng, David G. Beer, James P. Hamilton, and Bogdan C. Paun
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Male ,Cancer Research ,Pathology ,medicine.medical_specialty ,Esophageal Neoplasms ,Adenocarcinoma ,Biology ,medicine.disease_cause ,chemistry.chemical_compound ,Risk Factors ,Cell Line, Tumor ,medicine ,Humans ,Esophagus ,Promoter Regions, Genetic ,DNA Primers ,Base Sequence ,Reverse Transcriptase Polymerase Chain Reaction ,Esophageal disease ,Cancer ,Promoter ,Methylation ,DNA Methylation ,Middle Aged ,Cadherins ,medicine.disease ,medicine.anatomical_structure ,Oncology ,chemistry ,DNA methylation ,Azacitidine ,Cancer research ,Female ,Deoxycytidine ,Carcinogenesis - Abstract
Although the CDH13 gene has been shown to undergo epigenetic silencing by promoter methylation in many types of tumors, hypermethylation of this gene in Barrett's-associated esophageal adenocarcinogenesis has not been studied. Two hundred fifty-nine human esophageal tissues were therefore examined for CDH13 promoter hypermethylation by real-time methylation-specific PCR. CDH13 hypermethylation showed discriminative receiver-operator characteristic curve profiles, sharply demarcating esophageal adenocarcinoma (EAC) from esophageal squamous cell carcinoma (ESCC) and normal esophagus (NE) (p < 0.0001). CDH13 normalized methylation values (NMV) were significantly higher in Barrett's esophagus (BE), dysplastic BE (D) and EAC than in NE (p < 0.0000001). CDH13 hypermethylation frequency was 0% in NE but increased early during neoplastic progression, rising to 70% in BE, 77.5% in D and 76.1% in EAC. Both CDH13 hypermethylation frequency and its mean NMV were significantly higher in BE with than without accompanying EAC. In contrast, only 5 (19.2%) of 26 ESCCs exhibited CDH13 hypermethylation. Furthermore, both CDH13 hypermethylation frequency and its mean NMV were significantly higher in EAC than in ESCC, as well as in BE or D vs. ESCC. Interestingly, mean CDH13 NMV was significantly lower in short-segment than in long-segment BE, a known clinical risk factor for neoplastic progression. Similarly, BE segment length was significantly lower in specimens with unmethylated than with methylated CDH13 promoters. 5-aza-2'-deoxycytidine treatment of OE33 EAC and KYSE220 ESCC cells reduced CDH13 methylation and increased CDH13 mRNA expression. These findings suggest that hypermethylation of CDH13 is a common, tissue-specific event in human EAC, occurs early during BE-associated neoplastic progression, and correlates with known clinical neoplastic progression risk factors.
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- 2008
8. Pituitary Tumor-Transforming 1 Increases Cell Motility and Promotes Lymph Node Metastasis in Esophageal Squamous Cell Carcinoma
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Takatsugu Kan, Yutaka Shimada, Alexandru Olaru, Jian Yang, Tetsuo Ito, Stephen J. Meltzer, Fumiaki Sato, Yulan Cheng, Stefan David, Yuriko Mori, Zhe Jin, Rachana Agarwal, Bogdan C. Paun, John M. Abraham, and James P. Hamilton
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Adult ,Male ,Cancer Research ,Pathology ,medicine.medical_specialty ,Esophageal Neoplasms ,Mice, Nude ,Motility ,Biology ,Transfection ,Article ,Mice ,Cell Movement ,Carcinoma ,medicine ,Animals ,Humans ,Gastrointestinal cancer ,RNA, Small Interfering ,Lymph node ,Cells, Cultured ,Aged ,Oligonucleotide Array Sequence Analysis ,Aged, 80 and over ,Gene Expression Profiling ,Middle Aged ,medicine.disease ,Survival Analysis ,digestive system diseases ,Neoplasm Proteins ,Gene Expression Regulation, Neoplastic ,Securin ,Gene expression profiling ,medicine.anatomical_structure ,Oncology ,Cell culture ,Lymphatic Metastasis ,Carcinoma, Squamous Cell ,Immunohistochemistry ,Female ,Lymph Nodes - Abstract
Human pituitary tumor-transforming 1 (PTTG1)/securin is a putative oncoprotein that is overexpressed in various tumor types. However, the involvement of PTTG1 in gastrointestinal cancer development and progression remains unclear. In this study, we investigated the clinical significance and biological effects of PTTG1 in esophageal squamous cell carcinoma (ESCC). Immunohistochemical studies performed on 113 primary ESCC specimens revealed a high prevalence of PTTG1 overexpression (60.2%), which was significantly associated with lymph node metastasis (regional, P = 0.042; distant, P = 0.005), advanced tumor stage (P = 0.028), and poorer overall survival (P = 0.017, log-rank test; P = 0.044, Cox proportional hazard model). Eleven ESCC cell lines expressed PTTG1 protein at levels 2.4 to 6.6 times higher than those in normal esophageal epithelial cells (HEEpiC). PTTG1 protein expression was confined to the nucleus in HEEpiC cells but present in both the cytoplasm and nucleus in ESCC cells. Two small interfering RNAs (siRNA) inhibited PTTG1 mRNA and protein expression in three ESCC cell lines by 77% to 97%. In addition, PTTG1 down-regulation by these siRNAs significantly reduced cell motility in all three ESCC cell lines (P < 0.01) in vitro, as well as popliteal lymph node metastases of ESCC cells in nude mice (P = 0.020). Global gene expression profiling suggested that several members of the Ras and Rho gene families, including RRAS, RHOG, ARHGAP1, and ARHGADIA, represented potential downstream genes in the PTTG1 pathway. Taken together, these findings suggest that PTTG1 overexpression promotes cell motility and lymph node metastasis in ESCC patients, leading to poorer survival. Thus, PTTG1 constitutes a potential biomarker and therapeutic target in ESCCs with lymph node metastases. [Cancer Res 2008;68(9):3214–24]
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- 2008
9. Hypermethylation of the nel-like 1 gene is a common and early event and is associated with poor prognosis in early-stage esophageal adenocarcinoma
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D.G. Beer, Tetsuo Ito, Stephen J. Meltzer, Rachana Agarwal, Yuriko Mori, Alexandru Olaru, John M. Abraham, James P. Hamilton, Bogdan C. Paun, Jian Yang, Stefan David, Zhe Jin, Elizabeth A. Montgomery, Yulan Cheng, Fumiaki Sato, and Takatsugu Kan
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Cancer Research ,Time Factors ,Esophageal Neoplasms ,NELL1 ,Cytidine Triphosphate ,Nerve Tissue Proteins ,Biology ,medicine.disease_cause ,Loss of heterozygosity ,chemistry.chemical_compound ,Cell Line, Tumor ,Metaplasia ,Genetics ,medicine ,Humans ,RNA, Messenger ,Promoter Regions, Genetic ,Molecular Biology ,Survival rate ,Aged ,Neoplasm Staging ,Calcium-Binding Proteins ,Methylation ,DNA Methylation ,Middle Aged ,Prognosis ,digestive system diseases ,Survival Rate ,chemistry ,DNA methylation ,Azacitidine ,Cancer research ,Deoxycytidine ,medicine.symptom ,Carcinogenesis - Abstract
The nel-like1 (NELL1) gene maps to chromosome 11p15, which frequently undergoes loss of heterozygosity in esophageal adenocarcinoma (EAC). NELL1 promoter hypermethylation was examined by real-time methylation-specific polymerase chain reaction in 259 human esophageal tissues. Hypermethylation of this promoter showed highly discriminative receiver-operator characteristic curve profiles, clearly distinguishing esophageal squamous cell carcinoma (ESCC) and EAC from normal esophagus (NE) (P
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- 2007
10. A Genome-Wide Search Identifies Epigenetic Silencing of Somatostatin, Tachykinin-1, and 5 Other Genes in Colon Cancer
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Stephen J. Meltzer, Yuriko Mori, Takatsugu Kan, Bogdan C. Paun, Agnes T. Berki, Fumiaki Sato, Yulan Cheng, John M. Abraham, Zhe Jin, Carmit Mantzur, Kun Cai, Suna Wang, Tetsuo Ito, and James P. Hamilton
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Adult ,Male ,Candidate gene ,Tumor suppressor gene ,Colorectal cancer ,Bisulfite sequencing ,In Vitro Techniques ,Substance P ,Biology ,medicine ,Humans ,Gene Silencing ,Epigenetics ,Aged ,Aged, 80 and over ,Regulation of gene expression ,Hepatology ,Genome, Human ,Reverse Transcriptase Polymerase Chain Reaction ,Gastroenterology ,DNA, Neoplasm ,Methylation ,DNA Methylation ,Middle Aged ,Microarray Analysis ,medicine.disease ,Molecular biology ,Gene Expression Regulation, Neoplastic ,Colonic Neoplasms ,DNA methylation ,Female ,Somatostatin ,Microsatellite Repeats - Abstract
Background & Aims: Gene silencing via promoter hypermethylation is a central event in the pathogenesis of cancers. To identify novel methylation targets in colon cancer, we conducted a genome-wide, microarray-based, in silico, and epigenetic search. Methods: Complementary DNA microarray experiments were first performed to identify genes down-regulated in primary colon cancers and up-regulated in colon cancer cell lines after global DNA demethylation by 5-aza-2′-deoxycitidine. Candidate methylation targets were then identified by combining these microarray data with in silico genetic and functional searches. Candidate genes recognized by these searches were further investigated for promoter hypermethylation in colon cancer using methylation-specific polymerase chain reaction. Results: We identified 51 novel and 3 known candidate methylation targets. Subsequent epigenetic analysis revealed that primary colon cancers demonstrated frequent methylation of somatostatin (SST, 30 of 34 cases, 88%) and the substance P precursor gene tachykinin-1 (TAC1; 16 of 34 cases, 47%). TAC1 methylation intensity was significantly higher in Dukes A/B than in Dukes C/D cancers (P = .01). SST methylation intensity was significantly higher in low-level microsatellite instability (MSI-L) than in non-MSI-L cancers (P = .02). Methylation was associated with messenger RNA down-regulation for both SST and TAC1. Furthermore, we isolated 5 additional novel promoter methylation targets: NELL1, AKAP12, caveolin-1, endoglin, and MAL. Conclusions: These data strongly suggest that SST and TAC1 are involved in colon carcinogenesis. Further studies are now indicated to elucidate mechanisms underlying their involvement in colon cancer and their values as clinical biomarkers. NELL1, AKAP12, caveolin-1, endoglin, and MAL are also promising tumor suppressor gene candidates deserving of further study.
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- 2006
11. Mutational and LOH Analyses of the Chromosome 4q Region in Esophageal Adenocarcinoma
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John M. Abraham, Stephen J. Meltzer, Takatsugu Kan, Bogdan C. Paun, Yuriko Mori, Kun Cai, Fumiaki Sato, Andreea Olaru, Anca Sterian, Agnes T. Berki, Karsten Schulmann, Suna Wang, and James P. Hamilton
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Cancer Research ,Pathology ,medicine.medical_specialty ,F-Box-WD Repeat-Containing Protein 7 ,Esophageal Neoplasms ,Tumor suppressor gene ,Ubiquitin-Protein Ligases ,DNA Mutational Analysis ,Loss of Heterozygosity ,Cell Cycle Proteins ,Adenocarcinoma ,Biology ,medicine.disease_cause ,Loss of heterozygosity ,medicine ,Humans ,Genes, Tumor Suppressor ,Esophagus ,Mutation ,Esophageal disease ,F-Box Proteins ,Cancer ,Chromosome ,General Medicine ,medicine.disease ,DNA-Binding Proteins ,medicine.anatomical_structure ,Oncology ,Chromosomes, Human, Pair 4 ,Carcinogenesis ,Gene Deletion ,Transcription Factors - Abstract
Objective: Mortality due to esophageal adenocarcinoma has risen markedly, but the molecular mechanisms underlying this carcinogenesis are still incompletely understood. Findings from loss of heterozygosity (LOH) studies have suggested that the long arm of chromosome 4 might harbor tumor suppressor genes relevant to esophageal adenocarcinoma. Methods: We performed LOH analysis of 4q in esophageal adenocarcinomas. Regions of LOH were further evaluated by studying two candidate tumor suppressor genes, hCDC4 and CARF, located within them. Results: 54% of the adenocarcinomas examined showed allelic deletion. LOH was observed in 53, 40, 32, 38, and 27% of tumors at positions D4S1554 (the locus of CARF), D4S1572, D4S1548, D4S2934, and D4S3021, respectively. An area of allelic deletion (spanning 3 million bases) was identified at 4q31.1–3 in 37% of tumors. This region harbors a candidate tumor suppressor gene: hCDC4. However, sequencing of the coding regions of CARF and hCDC4 at 4q35 and 4q31, respectively, did not identify mutations. Conclusions: Our findings demonstrate frequent LOH in esophageal adenocarcinoma at several loci including a novel area of allelic deletion at 4q31.1–3. The results imply that mutational or other alterations at these loci may be involved in the pathogenesis of esophageal adenocarcinoma. Candidate tumor suppressor genes located within these regions merit further study.
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- 2006
12. Inactivation of p16, RUNX3, and HPP1 occurs early in Barrett's-associated neoplastic progression and predicts progression risk
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Stephen J. Meltzer, Karsten Schulmann, Yuriko Mori, Fumiaki Sato, Andreea Olaru, Ziding Feng, Yan Xu, James P. Hamilton, Suna Wang, Agnes Berki, Takatsugu Kan, Elena Deacu, Margaret S. Pepe, John M. Abraham, Wolff Schmiegel, Anca Sterian, Bruce D. Greenwald, David G. Beer, Mark J. Krasna, and Jing Yin
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Oncology ,Cancer Research ,medicine.medical_specialty ,Esophageal Neoplasms ,Adenocarcinoma ,Biology ,Polymerase Chain Reaction ,Barrett Esophagus ,Risk Factors ,Cell Line, Tumor ,Internal medicine ,Genetics ,medicine ,Humans ,RNA, Neoplasm ,Promoter Regions, Genetic ,Molecular Biology ,Cyclin-Dependent Kinase Inhibitor p16 ,Reverse Transcriptase Polymerase Chain Reaction ,Esophageal disease ,Proportional hazards model ,Membrane Proteins ,Cancer ,DNA, Neoplasm ,Odds ratio ,DNA Methylation ,Esophageal cancer ,medicine.disease ,Neoplasm Proteins ,DNA-Binding Proteins ,Core Binding Factor Alpha 3 Subunit ,Dysplasia ,Barrett's esophagus ,DNA methylation ,Immunology ,Disease Progression ,Transcription Factors - Abstract
Patients with Barrett's esophagus (BE) are at increased risk of developing esophageal adenocarcinoma (EAC). Clinical neoplastic progression risk factors, such as age and the length of the esophageal BE segment, have been identified. However, improved molecular biomarkers predicting increased progression risk are needed for improved risk assessment and stratification. Using real-time quantitative methylation-specific PCR, we screened 10 genes (HPP1, RUNX3, RIZ1, CRBP1, 3-OST-2, APC, TIMP3, p16, MGMT, p14) for promoter hypermethylation in 77 EAC, 93 BE, and 64 normal esophagus (NE) specimens. A subset of genes manifesting significant differences in methylation frequencies between BE and EAC was then analysed in 20 dysplastic specimens. All 10 genes except p14 were frequently methylated in EACs, with RUNX3, HPP1, CRBP1, RIZ1, and OST-2 representing novel methylation targets in EAC and/or BE. p16, RUNX3, and HPP1 displayed increasing methylation frequencies in BE vs EAC. Furthermore, these increases in methylation occurred early, at the interface between BE and low-grade dysplasia (LGD). To demonstrate the silencing effect of hypermethylation, we selected the EAC cells BIC1, in which the HPP1 promoter is natively methylated, and subjected them to 5-aza-2'-deoxycytidine (Aza-C) treatment. Real-time RT-PCR indicated increased HPP1 mRNA levels after 3 days of Aza-C treatment, as well as decreased levels of methylated HPP1 DNA. Hypermethylation of a subset of six genes (APC, TIMP3, CRBP1, p16, RUNX3, and HPP1) was then tested in a retrospective longitudinal study of 99 BE and nine LGD specimens obtained from 53 BE patients undergoing surveillance endoscopy. Only high-grade dysplasia (HGD) or EAC were defined as progression end points. Two patient groups were compared: eight progressors (P) and 45 nonprogressors (NP), using Cox proportional hazards models to determine the relative progression risks of age, BE segment length, and methylation events. Multivariate analyses revealed that only hypermethylation of p16 (odds ratio (OR) 1.74, 95% confidence interval (CI) 1.33-2.20), RUNX3 (OR 1.80, 95% CI 1.08-2.81), and HPP1 (OR 1.77, 95% CI 1.06-2.81) were independently associated with an increased risk of progression, whereas age, BE segment length, and hypermethylation of TIMP3, APC, or CRBP1 were not independent risk factors. In combined analyses, risk was detectable up to, but not earlier than, 2 years preceding neoplastic progression. Hypermethylation of p16, RUNX3, and HPP1 in BE or LGD may represent independent risk factors for the progression of BE to HGD or EAC. These findings have implications regarding risk stratification, early EAC detection, and the appropriate endoscopic surveillance interval for patients with BE.
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- 2005
13. The Clinical Significance of Aurora-A/STK15/BTAK Expression in Human Esophageal Squamous Cell Carcinoma
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Tetsuo Ito, Yutaka Shimada, Takatsugu Kan, Go Watanabe, Tomoyuki Okumura, Johji Inazawa, Eiji Tanaka, Yosuke Hashimoto, and Masayuki Imamura
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Male ,Cancer Research ,Pathology ,medicine.medical_specialty ,Esophageal Neoplasms ,Blotting, Western ,Cell ,macromolecular substances ,Protein Serine-Threonine Kinases ,Biology ,Metastasis ,Western blot ,Aurora Kinases ,Chromosome instability ,Tumor Cells, Cultured ,medicine ,Humans ,Aged ,Aurora Kinase A ,medicine.diagnostic_test ,Reverse Transcriptase Polymerase Chain Reaction ,Kinase ,Gene Expression Profiling ,Cancer ,Middle Aged ,Prognosis ,medicine.disease ,Epithelium ,Up-Regulation ,medicine.anatomical_structure ,Oncology ,Lymphatic Metastasis ,embryonic structures ,Carcinoma, Squamous Cell ,Immunohistochemistry ,Female ,biological phenomena, cell phenomena, and immunity - Abstract
Purpose: Aurora-A/STK15/BTAK (Aurora-A) encodes a Serine/Threonine kinase associated with chromosomal distribution, and its up-regulation induces chromosomal instability thereby leading to aneuploidy and cell transformation in several types of cancer. In this study, we investigated the role of Aurora-A in human esophageal squamous cell carcinoma (ESCC). Experimental Design: The expression levels of Aurora-A mRNA were compared in 33 ESCC tissues with that in corresponding normal esophageal epithelium by semiquantitative reverse transcription-PCR, and the distribution patterns and expression levels of Aurora-A protein were immunohistochemically investigated in the ESCC tumors of 142 patients. The results were then separately compared with the clinicopathologic findings of the patients, and the expression of Aurora-A was examined in nine ESCC cell lines and a normal esophageal epithelial cell line using Western blot analysis. Results: The up-regulation of Aurora-A mRNA was found in 30% (10 of 33) of the tumors by semiquantitative reverse transcription-PCR, and protein up-regulation was found in 53% (75 of 142) of the patients by immunohistochemistry. mRNA and protein up-regulation of Aurora-A were correlated with distant lymph node metastasis (P = 0.05 and P = 0.04, respectively), and patients with Aurora-A mRNA or protein up-regulation had a poorer prognosis (P = 0.003 and P = 0.0009, respectively). Furthermore, multivariate analysis revealed that up-regulation of the Aurora-A protein was an independent prognostic factor. In addition, Aurora-A expression in all ESCC cell lines was higher than that in a normal esophageal epithelial cell line. Conclusions: The up-regulation of Aurora-A expression may reflect the malignant behavior of ESCC and may prove useful information as a prognostic factor for ESCC patients.
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- 2005
14. Prediction of Lymph Node Metastasis with Use of Artificial Neural Networks Based on Gene Expression Profiles in Esophageal Squamous Cell Carcinoma
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Tetsuo Ito, Masato Maeda, Kan Kondo, Stephen J. Meltzer, Go Watanabe, Fumiaki Sato, Masayuki Imamura, Seiji Yamasaki, Yutaka Shimada, and Takatsugu Kan
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Adult ,Male ,Oncology ,Candidate gene ,medicine.medical_specialty ,Esophageal Neoplasms ,Microarray ,Text mining ,Internal medicine ,Carcinoma ,medicine ,Humans ,Lymph node ,Aged ,Oligonucleotide Array Sequence Analysis ,Aged, 80 and over ,Reverse Transcriptase Polymerase Chain Reaction ,business.industry ,Gene Expression Profiling ,Middle Aged ,Esophageal cancer ,Prognosis ,medicine.disease ,Gene expression profiling ,medicine.anatomical_structure ,Lymphatic Metastasis ,Significance analysis of microarrays ,Carcinoma, Squamous Cell ,Female ,Surgery ,Neural Networks, Computer ,business ,Forecasting - Abstract
Background: The aim of the study was (1) to detect candidate genes involved in lymph node metastasis in esophageal cancers and (2) to investigate whether we can estimate and predict occurrence of lymph node metastasis by analyzing artificial neural networks (ANNs) using these gene subsets. Methods: Twenty-eight primary esophageal squamous cell carcinomas were used. Gene expression profiles of all primary tumors were obtained by cDNA microarray. Lymph node metastasis–related genes were extracted with use of Significance Analysis of Microarrays (SAM). Predictive accuracy for lymph node metastasis was calculated by evaluation of 28 cases by ANNs with leave-one-out cross-n. The results were compared with those of other analyses such as clustering or predictive scoring (LMS). Results: Our ANN model could predict lymph node metastasis most accurately with 60 clones. The highest predictive accuracy for lymph node metastasis by ANN was 10 of 13 (77%) in newly added cases that were not used for gene selection by SAM and 24 of 28 (86%) in all cases (sensitivity: 15/17, 88%; specificity: 9/11, 82%). Predictive accuracy of LMS was 9 of 13 (69%) in newly added cases and 24 of 28 (86%) in all cases (sensitivity: 17/17, 100%; specificity: 7/11, 67%). It was difficult to extract useful information for the prediction of lymph node metastasis by clustering analysis. Conclusions: ANN had superior potential in comparison with other methods of analysis for the prediction of lymph node metastasis. This systematic analysis combining SAM with ANN was very useful for the prediction of lymph node metastasis in esophageal cancers and could be applied clinically in the near future.
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- 2004
15. Activin Type II Receptor Restoration in ACVR2 -Deficient Colon Cancer Cells Induces Transforming Growth Factor-β Response Pathway Genes
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Takatsugu Kan, Fumiaki Sato, Stephen J. Meltzer, Yuriko Mori, Andreea Olaru, Yan Xu, Karsten Schulmann, Elena Deacu, Jing Yin, Suna Wang, Anca Sterian, Agnes T. Berki, and John M. Abraham
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Cancer Research ,medicine.medical_specialty ,Tumor suppressor gene ,Activin Receptors, Type II ,Smad2 Protein ,SMAD ,Protein Serine-Threonine Kinases ,Biology ,Mothers against decapentaplegic homolog 3 ,Transforming Growth Factor beta ,Cell Line, Tumor ,Internal medicine ,TGF beta signaling pathway ,medicine ,Humans ,Phosphorylation ,Activin type 2 receptors ,Receptor, Transforming Growth Factor-beta Type II ,Cell biology ,DNA-Binding Proteins ,Gene Expression Regulation, Neoplastic ,Endocrinology ,Oncology ,Colonic Neoplasms ,Trans-Activators ,Signal transduction ,Receptors, Transforming Growth Factor beta ,Cell Division ,ACVR2B ,Signal Transduction ,Transforming growth factor - Abstract
The activin type II receptor (ACVR2) gene is a putative tumor suppressor gene that is frequently mutated in microsatellite-unstable colon cancers (MSI-H colon cancers). ACVR2 is a member of the transforming growth factor (TGF)-β type II receptor (TGFBR2) family and controls cell growth and differentiation. SMAD proteins are major intracellular effectors shared by ACVR2 and TGFBR2 signaling; however, additional shared effector mechanisms remain to be explored. To discover novel mechanisms transmitting the ACVR2 signal, we restored ACVR2 function by transfecting wild-type ACVR2 (wt-ACVR2) into a MSI-H colon cancer cell line carrying an ACVR2 frameshift mutation. The effect of ACVR2 restoration on cell growth, SMAD phosphorylation, and global molecular phenotype was then evaluated. Decreased cell growth was observed in wt-ACVR2 transfectants relative to ACVR2-deficient vector-transfected controls. Western blotting revealed higher expression of phosphorylated SMAD2 in wt-ACVR2 transfectants versus controls, suggesting cells deficient in ACVR2 had impaired SMAD signaling. Microarray-based differential expression analysis revealed substantial ACVR2-induced overexpression of genes implicated in the control of cell growth and tumorigenesis, including the activator protein (AP)-1 complex genes JUND, JUN, and FOSB, as well as the small GTPase signal transduction family members, RHOB, ARHE, and ARHGDIA. Overexpression of these genes is shared with TGFBR2 activation. This observed similarity between the activin and TGF-β signaling systems suggests that activin may serve as an alternative activator of TGF-β effectors, including SMADs, and that frameshift mutation of ACVR2 may contribute to MSI-H colon tumorigenesis via disruption of alternate TGF-β effector pathways.
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- 2004
16. Validity of intraoperative pathological diagnosis of paratracheal lymph node as a strategy for selection of patients for cervical lymph node dissection during esophagectomy
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Masato Maeda, Yosuke Hashimoto, Zhigang Li, Junichi Kaganoi, Shiro Nagatani, Yutaka Shimada, Masayuki Imamura, Fumiaki Sato, Go Watanabe, and Takatsugu Kan
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Male ,medicine.medical_specialty ,Esophageal Neoplasms ,medicine.medical_treatment ,Intraoperative Period ,medicine ,Paratracheal ,Humans ,Lymph node ,Aged ,Retrospective Studies ,business.industry ,Patient Selection ,Paratracheal lymph nodes ,Micrometastasis ,Gastroenterology ,Reproducibility of Results ,General Medicine ,Middle Aged ,Esophageal cancer ,medicine.disease ,Surgery ,Esophagectomy ,Trachea ,Dissection ,medicine.anatomical_structure ,Lymphatic Metastasis ,Lymph Node Excision ,Female ,business ,Neck - Abstract
The aim of this paper is to examine whether intraoperative examination of paratracheal nodes can indicate cervical node dissection and whether this approach is valid. From 1988 to 1997, 76 patients with thoracic esophageal squamous cell carcinoma received esophagectomies with and without cervical lymph node (LN) dissection based on the results of intraoperative pathological diagnosis from selective checking of paratracheal LN. We retrospectively examined the outcomes for the patients and the micro metastasis in the dissected lymph node using cytokeratin staining. Three of the seven patients with cervical LN dissection were detected as having cervical LN metastasis by postoperative hematoxylin-eosin or cytokeratin staining. Five (7%) of the 69 patients without cervical LN dissection had cervical LN recurrence after the operation. Four of the seven patients who were diagnosed as having metastasis or micro metastasis in paratracheal LN by postoperative examination had cervical LN recurrence after the operation. In conclusion, the esophagectomy with and without cervical LN dissection for thoracic esophageal squamous cell carcinoma based on the results of intraoperative pathological diagnosis from selective checking of paratracheal LN was not fully acceptable. The reliability of intraoperative pathological diagnosis of selective checking may improve by increasing the number of checked LN and the detection of micro metastasis.
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- 2003
17. [Two cases of curative resection by laparoscopic surgery following preoperative chemotherapy with bevacizumab for locally advanced colon cancer]
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Masazumi, Sakaguchi, Takatsugu, Kan, Michihiko, Tsubono, and Eiji, Kii
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Bevacizumab ,Male ,Sigmoid Neoplasms ,Humans ,Female ,Laparoscopy ,Neoplasm Invasiveness ,Middle Aged ,Antibodies, Monoclonal, Humanized ,Combined Modality Therapy ,Neoplasm Staging - Abstract
Here we report 2 cases of curative resection following preoperative chemotherapy with bevacizumab for locally advanced colon cancer. Case 1 was a 62-year-old man admitted with constipation, abdominal distention, and abdominal pain. An abdominal computed tomography(CT)scan revealed an obstructive tumor of the sigmoid colon with invasion into the bladder. A diverting colostomy was performed, and chemotherapy with mFOLFOX6(infusional 5-fluorouracil/Leucovorin+ oxaliplatin) plus bevacizumab was initiated. The tumor shrunk markedly after 6 courses of this treatment. Thereafter, laparoscopy- assisted sigmoidectomy was successfully performed. Case 2 was a 61-year-old woman admitted with diarrhea, abdominal pain, and fever. An abdominal CT scan revealed an obstructive tumor of the sigmoid colon with invasion into the ileum, uterus and retroperitoneum. A diverting colostomy was performed, and chemotherapy with XELOX(capecitabine+ oxaliplatin)plus bevacizumab was initiated. The tumor shrunk markedly after 6 courses of this treatment. Thereafter, laparoscopy- assisted sigmoidectomy was successfully performed. Both cases demonstrated partial clinical responses to chemotherapy; thus, curative resection surgeries were performed. There were no perioperative complications. Therefore, we conclude that oxaliplatin-based chemotherapy plus bevacizumab and laparoscopic resection could be very effective for locally advanced colon cancer.
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- 2014
18. MicroRNA-192 and -215 are upregulated in human gastric cancer in vivo and suppress ALCAM expression in vitro
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Mark D. Duncan, Stephen J. Meltzer, Jian Yang, Takatsugu Kan, Yuriko Mori, Florin M. Selaru, John W. Harmon, Rachana Agarwal, Elizabeth A. Montgomery, Yulan Cheng, James P. Hamilton, Stefan David, Zhe Jin, Alexandru Olaru, and John M. Abraham
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Fetal Proteins ,Cancer Research ,Cell Adhesion Molecules, Neuronal ,Apoptosis ,Biology ,Article ,Downregulation and upregulation ,In vivo ,Antigens, CD ,Stomach Neoplasms ,Cell Line, Tumor ,microRNA ,Genetics ,medicine ,Humans ,Molecular Biology ,ALCAM ,Cell Proliferation ,Oligonucleotide Array Sequence Analysis ,Cell growth ,Reverse Transcriptase Polymerase Chain Reaction ,Cell Cycle ,Cancer ,Cell cycle ,medicine.disease ,In vitro ,Cell biology ,Up-Regulation ,Gene Expression Regulation, Neoplastic ,MicroRNAs - Abstract
The dismal outcome of gastric cancer patients highlights the need for diagnostic biomarkers and effective therapeutic targets, such as microRNAs. We sought to discover microRNAs involved in gastric cancer, and to elucidate their downstream target mechanisms. Both cultured gastric epithelial cells (HFE145 and NCI-N87) and primary human gastric tissues (31 non-neoplastic stomach (NS) and 25 gastric carcinomas (GC)) were studied. MicroRNA microarrays and quantitative RT–PCR were applied to discover and verify differentially expressed microRNAs. in vitro cell migration and invasion, cell proliferation, cell cycle and apoptosis assays were executed to elucidate biological effects of micro-RNA-192 and -215. Western blotting and luciferase assays were performed to confirm direct messenger RNA targeting by microRNA-192 and -215. MicroRNA microarray analyses revealed that 25 and 20 microRNAs were upregulated and downregulated in GC vs NS, respectively. Expression levels of both microRNA-192 and -215 were significantly higher in GC than in NS (P < 0.05). Luciferase assays suggested that microRNA-215 inhibits activated leukocyte cell adhesion molecule (ALCAM) expression at the posttranscriptional level. In addition, expression levels of ALCAM were significantly lower in GC than in NS. Mimics and inhibitors, respectively, of microRNA-192 or -215 exerted no effect on cell cycle or apoptosis in the immortalized normal gastric cell line HFE145 or the gastric cancer cell line NCI-N87. However, mimics of microRNA-192 or -215 significantly increased growth rates in HFE145 cells, whereas inhibitors of microRNA-192 or -215 caused significant decreases in growth rates in NCI-N87 cells. ALCAM knockdown by an ALCAM-specific siRNA significantly increased cell growth in HFE145 cells. Both transfection of mimics of microRNA-192 or -215 and ALCAM knockdown by an ALCAM-specific siRNA significantly increased the migration of HFE145 cells. In conclusion, in gastric cancer, both microRNA-192 and -215 are overexpressed in vivo and exert cell growth and migration-promoting effects in vitro, thus representing potential microRNAs with a role in cancer in the human stomach.
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- 2010
19. Polo-like kinase 1 regulates cell proliferation and is targeted by miR-593* in esophageal cancer
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Zhe Jin, Stefan David, Tetsuo Ito, John M. Abraham, Bogdan C. Paun, Kazuharu Shimizu, Jian Yang, Stephen J. Meltzer, Yuriko Mori, Florin M. Selaru, Alexandru Olaru, Yutaka Shimada, Rachana Agarwal, Yulan Cheng, Fumiaki Sato, Nobutoshi Matsumura, James P. Hamilton, and Takatsugu Kan
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Adult ,Male ,Cancer Research ,Esophageal Neoplasms ,RNA Stability ,Cell ,Mice, Nude ,Cell Cycle Proteins ,Biology ,Protein Serine-Threonine Kinases ,Article ,Mice ,Cell Line, Tumor ,Proto-Oncogene Proteins ,microRNA ,medicine ,Animals ,Humans ,Luciferase ,Gene Silencing ,RNA, Messenger ,3' Untranslated Regions ,Aged ,Cell Proliferation ,Aged, 80 and over ,Gene knockdown ,Base Sequence ,Cell growth ,Cell cycle ,Middle Aged ,Molecular biology ,Gene Expression Regulation, Neoplastic ,MicroRNAs ,medicine.anatomical_structure ,Oncology ,Cell culture ,Apoptosis ,Cancer research - Abstract
Polo-like kinase 1 (PLK1) is overexpressed in various human cancers. However, the biological functions and the post-transcriptional regulations of PLK1 in esophageal cancer (EC) are still unknown. The purposes of our study are to determine whether PLK1 can be a molecular target of EC therapy and to identify a microRNA (miRNA) targeting PLK1. We performed loss-of-function and gain-of-function experiments regarding cell proliferation, cell cycle, apoptosis, in vivo tumor formation and luciferase reporter assays, using siRNAs against PLK1 and miRNA. PLK1 protein was expressed in all 11 EC cell lines, but not in normal esophageal epithelial cells (HEEpiC). Knockdown of PLK1 in EC cells induced G2/M arrest (p < 0.001) in cell cycle assay and reduced cell proliferation (p = 0.019) and tumor formation ability in vivo (p < 0.0001). MiR-593*, identified as a miRNA targeting PLK1 by a database search, was less expressed especially in six EC cell lines than HEEpiC cells. Moreover, miR-593* expression level was inversely correlated with PLK1 mRNA level in 48 clinical tissue specimens of EC (p = 0.006). Introduction of synthetic miR-593* suppressed PLK1 expression by 69-73%, reduced cell proliferation (p = 0.008) and increased cell proportion of G2/M phase (p = 0.01) in HSA/c (an EC cells), whereas a miR-593* inhibitor upregulated PLK1 expression by 11-55%. Additionally, luciferase assay demonstrated that miR-593* interacted two binding sites in the PLK1 3'-UTR and reduced 56.8-71.5% of luciferase activity by degrading luciferase mRNA in HSA/c cells. In conclusion, PLK1 is post-transcriptionally regulated by miR-593* and could be a promising molecular target for EC treatment.
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- 2010
20. A Multicenter, Double-Blinded Validation Study of Methylation Biomarkers for Progression Prediction in Barrett's Esophagus
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Fumiaki Sato, Tetsuo Ito, Jian Yang, Ziding Feng, Mark A. Nelson, Bogdan C. Paun, Stephen J. Meltzer, James P. Hamilton, Alexandru Olaru, Yuriko Mori, Kay Washington, Florin M. Selaru, Jean Wang, Yulan Cheng, Richard E. Sampliner, Michael B. Wallace, Marcia I. Canto, Rachana Agarwal, Paul D. Wagner, Zhe Jin, Nicholas J. Shaheen, Kenneth K. Wang, Yvonne Romero, Yingye Zheng, Takatsugu Kan, Herbert C. Wolfsen, Stefan David, Achyut K. Bhattacharyya, Wen Gu, and John M. Abraham
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Cancer Research ,medicine.medical_specialty ,Esophageal Neoplasms ,Population ,Logistic regression ,Polymerase Chain Reaction ,Risk Assessment ,Gastroenterology ,Article ,Barrett Esophagus ,Double-Blind Method ,Predictive Value of Tests ,Internal medicine ,medicine ,Humans ,Esophagus ,Promoter Regions, Genetic ,education ,Cyclin-Dependent Kinase Inhibitor p16 ,education.field_of_study ,medicine.diagnostic_test ,business.industry ,Esophageal disease ,Age Factors ,Membrane Proteins ,Reproducibility of Results ,Endoscopy ,medicine.disease ,Neoplasm Proteins ,Gene Expression Regulation, Neoplastic ,Core Binding Factor Alpha 3 Subunit ,medicine.anatomical_structure ,Gene Expression Regulation ,ROC Curve ,Oncology ,Predictive value of tests ,Barrett's esophagus ,Disease Progression ,Biomarker (medicine) ,business ,Biomarkers - Abstract
Esophageal adenocarcinoma risk in Barrett's esophagus (BE) is increased 30- to 125-fold versus the general population. Among all BE patients, however, neoplastic progression occurs only once per 200 patient-years. Molecular biomarkers are therefore needed to risk-stratify patients for more efficient surveillance endoscopy and to improve the early detection of progression. We therefore performed a retrospective, multicenter, double-blinded validation study of eight BE progression prediction methylation biomarkers. Progression or nonprogression were determined at 2 years (tier 1) and 4 years (tier 2). Methylation was assayed in 145 nonprogressors and 50 progressors using real-time quantitative methylation-specific PCR. Progressors were significantly older than nonprogressors (70.6 versus 62.5 years; P < 0.001). We evaluated a linear combination of the eight markers, using coefficients from a multivariate logistic regression analysis. Areas under the ROC curve (AUC) were high in the 2-year, 4-year, and combined data models (0.843, 0.829, and 0.840; P < 0.001
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- 2009
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21. Generation of small 32P-labeled peptides as a potential approach to colorectal cancer therapy
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Takatsugu Kan, James P. Hamilton, Stephen J. Meltzer, John M. Abraham, Yuriko Mori, Florin M. Selaru, Tetsuo Ito, Rachana Agarwal, Stefan David, Alexandru Olaru, Jian Yang, Zhe Jin, Yulan Cheng, and Bogdan C. Paun
- Subjects
Gastroenterology and Hepatology/Gastrointestinal Cancers ,Colorectal cancer ,Genetics and Genomics/Pharmacogenomics ,lcsh:Medicine ,Oncology/Gastrointestinal Cancers ,Adenocarcinoma ,Bioinformatics ,03 medical and health sciences ,0302 clinical medicine ,Cell Line, Tumor ,Humans ,Medicine ,Phosphorylation ,lcsh:Science ,Molecular Biology ,Biotechnology/Small Molecule Chemistry ,030304 developmental biology ,CD20 ,chemistry.chemical_classification ,0303 health sciences ,Multidisciplinary ,biology ,business.industry ,lcsh:R ,Cancer ,medicine.disease ,3. Good health ,Lymphoma ,Amino acid ,chemistry ,Cell culture ,030220 oncology & carcinogenesis ,biology.protein ,Cancer research ,Autoradiography ,Electrophoresis, Polyacrylamide Gel ,lcsh:Q ,Biochemistry/Drug Discovery ,Antibody ,Colorectal Neoplasms ,Peptides ,Pharmacology/Personalized Medicine ,business ,Phosphorus Radioisotopes ,Research Article - Abstract
Cancers have been revealed to be extremely heterogenous in terms of the frequency and types of mutations present in cells from different malignant tumors. Thus, it is likely that uniform clinical treatment is not optimal for all patients, and that the development of individualized therapeutic regimens may be beneficial. We describe the generation of multiple, unique small peptides nine to thirty-four amino acids in length which, when labeled with the radioisotope (32)P, bind with vastly differing efficiencies to cell lines derived from different colon adenocarcinomas. In addition, the most effective of these peptides permanently transfers the (32)P radioisotope to colorectal cancer cellular proteins within two hours at a rate that is more than 150 times higher than in cell lines derived from other cancers or from the normal tissues tested. Currently, the only two FDA-approved radioimmunotherapeutic agents in use both employ antibodies directed against the B cell marker CD20 for the treatment of non-Hodgkin's lymphoma. By using the method described herein, large numbers of different (32)P-labeled peptides can be readily produced and assayed against a broad spectrum of cancer types. This report proposes the development and use of (32)P-labeled peptides as potential individualized peptide-binding therapies for the treatment of colon adenocarcinoma patients.
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- 2008
22. Hypermethylation of the AKAP12 promoter is a biomarker of Barrett's-associated esophageal neoplastic progression
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Tetsuo Ito, James P. Hamilton, Rachana Agarwal, John M. Abraham, Jian Yang, Yulan Cheng, David G. Beer, Stefan David, Bogdan C. Paun, Stephen J. Meltzer, Yuriko Mori, Takatsugu Kan, Fumiaki Sato, Zhe Jin, and Alexandru Olaru
- Subjects
medicine.medical_specialty ,Esophageal Neoplasms ,Epidemiology ,A Kinase Anchor Proteins ,Cell Cycle Proteins ,Biology ,Adenocarcinoma ,Gastroenterology ,Barrett Esophagus ,Internal medicine ,Metaplasia ,medicine ,Carcinoma ,Biomarkers, Tumor ,Tumor Cells, Cultured ,Humans ,RNA, Messenger ,Esophagus ,Promoter Regions, Genetic ,Methylation ,Esophageal cancer ,DNA Methylation ,medicine.disease ,digestive system diseases ,medicine.anatomical_structure ,Cell Transformation, Neoplastic ,Oncology ,Tumor progression ,Case-Control Studies ,DNA methylation ,Carcinoma, Squamous Cell ,Disease Progression ,Biomarker (medicine) ,medicine.symptom - Abstract
The A-kinase anchoring protein 12 (AKAP12) is a kinase scaffold protein with known tumor suppressor activity. Recently, AKAP12 promoter hypermethylation was reported in gastric and colorectal cancers. We examined AKAP12 promoter hypermethylation using real-time methylation-specific PCR in 259 human esophageal tissues. AKAP12 hypermethylation showed highly discriminative receiver-operator characteristic (ROC) curve profiles, clearly distinguishing esophageal adenocarcinoma (EAC) from esophageal squamous cell carcinoma and normal esophagus (P < 0.0001). AKAP12-normalized methylation values were significantly higher in Barrett's metaplasia (BE), dysplastic Barrett's, and EAC than in normal esophagus (P < 0.0000001). AKAP12 hypermethylation frequency was zero in normal esophagus but increased early during neoplastic progression, to 38.9% in BE from patients with Barrett's alone, 52.5% in dysplastic Barrett's metaplasia, and 52.2% in EAC. AKAP12 hypermethylation levels were significantly higher in normal esophageal epithelia from patients with EAC (mean = 0.00082) than in normal esophagi from patients without Barrett's or esophageal cancer (mean = 0.00007; P = 0.006). There was a significant correlation between AKAP12 hypermethylation and BE segment length, a known clinical neoplastic progression risk factor. In contrast, only 2 (7.7%) of 26 esophageal squamous cell carcinomas exhibited AKAP12 hypermethylation. Treatment of BIC and OE33 EAC cells with 5-aza-2'-deoxycytidine reduced AKAP12 methylation and increased AKAP12 mRNA expression. AKAP12 mRNA levels in EACs with unmethylated AKAP12 (mean = 0.1663) were higher than in EACs with methylated AKAP12 (mean = 0.0668). We conclude that promoter hypermethylation of AKAP12 is a common, tissue-specific event in human EAC, occurs early during Barrett's-associated esophageal neoplastic progression, and is a potential biomarker for the early detection of EAC. (Cancer Epidemiol Biomarkers Prev 2008;17(1):111–7)
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- 2008
23. Hypermethylation of tachykinin-1 is a potential biomarker in human esophageal cancer
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Tetsuo Ito, Bogdan C. Paun, Rachana Agarwal, John M. Abraham, Jian Yang, David G. Beer, Yulan Cheng, James P. Hamilton, Alexandru Olaru, Stephen J. Meltzer, Yuriko Mori, Takatsugu Kan, Carmit Mantzur, Fumiaki Sato, Zhe Jin, Suna Wang, and Stefan David
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Risk ,Cancer Research ,medicine.medical_specialty ,Esophageal Neoplasms ,Biology ,Gastroenterology ,Methylation ,Sensitivity and Specificity ,Cell Line ,Metaplasia ,Internal medicine ,Tachykinins ,medicine ,Carcinoma ,Biomarkers, Tumor ,Humans ,Neoplastic transformation ,Esophagus ,Promoter Regions, Genetic ,DNA ,Esophageal cancer ,DNA Methylation ,medicine.disease ,Prognosis ,digestive system diseases ,medicine.anatomical_structure ,Cell Transformation, Neoplastic ,Oncology ,DNA methylation ,Biomarker (medicine) ,RNA ,medicine.symptom ,Biomarkers - Abstract
Purpose: Our aim was to investigate whether and at what stage hypermethylation of the tachykinin-1 (TAC1) gene is associated with human esophageal neoplastic transformation.Experimental Design: TAC1 promoter hypermethylation was examined by real-time methylation-specific PCR in 258 human esophageal specimens and 126 plasma samples from patients or tissues at various stages of neoplastic evolution.Results: TAC1 hypermethylation in tissue samples showed highly discriminative receiver-operator characteristic curve profiles, clearly distinguishing esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma (EAC) from normal esophagus (P < 0.0001). Both frequencies and normalized methylation values of TAC1 tissue methylation were significantly higher in Barrett's metaplasia (BE), dysplastic Barrett's esophagus, EAC, and ESCC than in normal esophagus (P < 0.01). The frequency of TAC1 hypermethylation increased dramatically and early during neoplastic progression, from 7.5% in normal esophagus to 55.6% in BE from patients with Barrett's metaplasia alone, 57.5% in dysplastic Barrett's esophagus, and 61.2% in EAC. There was a significant relationship between TAC1 hypermethylation and BE segment length, a known clinical risk factor for neoplastic progression. Twelve (50%) of 24 ESCC exhibited TAC1 hypermethylation. Overall patient survival correlated significantly with TAC1 methylation status in ESCC patients (mean survival, 22 versus 110 months; P = 0.0102, log-rank test), but not in EAC patients. Both mean normalized methylation values and frequency of TAC1 hypermethylation in plasma samples were significantly higher in EAC patients than in control subjects. Treatment of KYSE220 ESCC and BIC EAC cells with 5-aza-2′-deoxycytidine reduced TAC1 methylation and increased TAC1 mRNA expression.Conclusions: TAC1 promoter hypermethylation is a common event in both major histologic types of human esophageal carcinoma, occurs early, correlates with other progression risk factors in esophageal adenocarcinogenesis, and is a tissue biomarker of a poor prognosis in ESCC. Circulating methylated TAC1 promoter DNA also offers potential as a biomarker for the diagnosis of EAC.
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- 2007
24. Reprimo methylation is a potential biomarker of Barrett's-Associated esophageal neoplastic progression
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Jian Yang, Tetsuo Ito, James P. Hamilton, Stephen J. Meltzer, Bruce D. Greenwald, Yuriko Mori, Fumiaki Sato, Takatsugu Kan, John M. Abraham, Suna Wang, Bogdan C. Paun, Yulan Cheng, and Zhe Jin
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Genetic Markers ,Male ,Cancer Research ,Pathology ,medicine.medical_specialty ,Esophageal Neoplasms ,Cell Cycle Proteins ,Biology ,Adenocarcinoma ,Decitabine ,Esophageal Diseases ,Polymerase Chain Reaction ,chemistry.chemical_compound ,Barrett Esophagus ,Cell Line, Tumor ,medicine ,Carcinoma ,Humans ,Esophagus ,Aged ,Glycoproteins ,Reprimo ,Methylation ,Esophageal cancer ,DNA Methylation ,Middle Aged ,medicine.disease ,digestive system diseases ,Demethylating agent ,medicine.anatomical_structure ,Oncology ,chemistry ,Dysplasia ,DNA methylation ,Cancer research ,Azacitidine ,Carcinoma, Squamous Cell ,Disease Progression ,Female ,Precancerous Conditions - Abstract
Purpose: Reprimo, a candidate tumor-suppressor gene, regulates p53-mediated cell cycle arrest at G2 phase, and tumor-suppressor gene methylation is involved in the pathogenesis and progression of esophageal cancer. Our aim was to determine whether and at what phase of neoplastic progression Reprimo methylation occurs in Barrett's adenocarcinogenesis, as well as its columnar or squamous cell-type specificity. We also sought to determine whether Reprimo expression could be restored in vitro by the demethylating agent 5-aza-deoxycytidine (5AzaC). Experimental Design: Quantitative methylation-specific PCR for Reprimo was done using an ABI7700 (Taqman) apparatus on 175 endoscopic biopsy specimens. In addition, reverse transcription-PCR and quantitative methylation-specific PCR were done on esophageal carcinoma cells before and after treatment with 5AzaC. Results: In Barrett's esophagus (BE; P = 0.001), high-grade dysplasia (HGD; P = 0.001), and esophageal adenocarcinoma (EAC; P = 0.00003), the level and frequency of Reprimo methylation were significantly higher than in normal esophagus (NE). There was no statistically significant difference between BE and EAC, HGD and EAC, or NE and esophageal squamous cell carcinoma (ESCC). Reprimo methylation occurred in 0 of 19 NE samples, 6 (13%) of 45 ESCC, 9 (36%) of 25 BE, 7 (64%) of 11 HGD, and 47 (63%) of 75 EAC. Analysis of Reprimo methylation in EAC versus NE revealed an area under the receiver-operator characteristic curve of 0.812 (P < 0.00001; 95% confidence interval, 0.73-0.90). In vitro 5AzaC treatment of OE33 EAC cells reduced Reprimo methylation and increased Reprimo expression. Conclusions: Reprimo methylation occurs significantly more frequently in BE, HGD, and EAC than in NE or ESCC, suggesting that this epigenetic alteration is a specialized columnar, cell-specific early event with potential as a biomarker for the early detection of esophageal neoplasia.
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- 2006
25. Promoter methylation and response to chemotherapy and radiation in esophageal cancer
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Tetsuo Ito, Takatsugu Kan, Carmit Mantzur, Bogdan C. Paun, Mohan Suntharalingam, Fumiaki Sato, Zhe Jin, Stephen J. Meltzer, James P. Hamilton, Yuriko Mori, Martin J. Edelman, Mark J. Krasna, Austin Doyle, John M. Abraham, Suna Wang, Bruce D. Greenwald, and Agnes T. Berki
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Oncology ,Adult ,Male ,medicine.medical_specialty ,Pathology ,Esophageal Neoplasms ,Antineoplastic Agents ,Adenocarcinoma ,Polymerase Chain Reaction ,symbols.namesake ,Internal medicine ,CHFR ,Carcinoma ,Medicine ,Humans ,Genes, Tumor Suppressor ,Gene Silencing ,Promoter Regions, Genetic ,Fisher's exact test ,Aged ,Aged, 80 and over ,Reprimo ,Hepatology ,business.industry ,Gastroenterology ,Methylation ,Esophageal cancer ,DNA Methylation ,Middle Aged ,medicine.disease ,Combined Modality Therapy ,Treatment Outcome ,DNA methylation ,symbols ,Carcinoma, Squamous Cell ,Biomarker (medicine) ,Female ,business - Abstract
Background & Aims: Multiple studies have shown that promoter methylation of tumor suppressor genes underlies esophageal carcinogenesis. Hypothetically, methylation resulting in tumor suppressor gene inactivation might result in tumors that are unresponsive to chemotherapy and radiation. Accordingly, our aim was to find methylation markers that could be used to predict response to chemoradiation. Methods: Tumor specimens were obtained before treatment from 35 patients enrolled in a uniform chemoradiation treatment protocol. Methylation-specific quantitative polymerase chain reaction was performed on all samples. Pathology reports from esophagectomy specimens were used to define response to treatment. Results: Thirteen (37%) of 35 patients were responders, and 22 (63%) of 35 patients were nonresponders. The number of methylated genes per patient was significantly lower in responders than in nonresponders (1.4 vs 2.4 genes per patient; Student t test, P = .026). The combined mean level of promoter methylation of p16, Reprimo, p57, p73, RUNX-3, CHFR, MGMT, TIMP-3, and HPP1 was also lower in responders than in nonresponders (Student t test, P = .003; Mann-Whitney test, P = .001). The frequency (15% of responders vs 64% of nonresponders; Fisher exact test, P = .01) and level (0.078 in responders vs 0.313 in nonresponders; Mann-Whitney test, P = .037) of Reprimo methylation was significantly lower in responders than in nonresponders. Conclusions: Reprimo methylation occurred at significantly lower levels and less frequently in chemoradioresponsive than in nonresponsive esophageal cancer patients, suggesting potential clinical application of this single-gene biomarker in defining prognosis and management. In addition, increased methylation of a 9-gene panel correlated significantly with poor responsiveness to chemoradiation.
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- 2006
26. Transcriptional profiling suggests that Barrett's metaplasia is an early intermediate stage in esophageal adenocarcinogenesis
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Bogdan C. Paun, M. M. Yu, Suna Wang, Yulan Cheng, Karsten Schulmann, John M. Abraham, Zhe Jin, Carmit Mantzur, Stephen J. Meltzer, James P. Hamilton, Yuriko Mori, Fumiaki Sato, Agnes T. Berki, Jing Yin, M. Zhan, H. Li, Alexandru Olaru, Bruce D. Greenwald, Tetsuo Ito, Yan Xu, and Takatsugu Kan
- Subjects
Cancer Research ,Microarray ,Esophageal Neoplasms ,Transcription, Genetic ,Biology ,Adenocarcinoma ,Barrett Esophagus ,Metaplasia ,Genetics ,medicine ,Biomarkers, Tumor ,Humans ,Molecular Biology ,Neoplasm Staging ,Oligonucleotide Array Sequence Analysis ,Esophageal disease ,Microarray analysis techniques ,Gene Expression Profiling ,Cancer ,Anatomy ,medicine.disease ,Gene expression profiling ,Cell Transformation, Neoplastic ,Barrett's esophagus ,Cancer research ,medicine.symptom - Abstract
To investigate the relationship between Barrett's esophagus (BE) and esophageal adenocarcinoma (EAC), we determined gene expression profiles of discrete pathological stages of esophageal neoplasia using a sequence-verified human cDNA microarray. Fifty one RNAs, comprising 24 normal esophagi (NE), 18 BEs, and nine EACs were hybridized to cDNA microarrays. Five statistical analyses were used for the data analysis. Genes showing significantly different expression levels among the three sample groups were identified. Genes were grouped into functional categories based on the Gene Ontology Consortium. Surprisingly, the expression pattern of BE was significantly more similar to EAC than to NE, notwithstanding the known histopathologic differences between BE and EAC. The pattern of NE was clearly distinct from that of EAC. Thirty-six genes were the most differentially modulated, according to these microarray data, in BE-associated neoplastic progression. Twelve genes were significantly differentially expressed in cancer-associated BE's plus EAC (as a single combined tissue group) vs noncancer-associated BE's. These genes represent potential biomarkers to diagnose EAC at its early stages. Our results demonstrate that molecular events at the transcriptional level in BE are remarkably similar to BE's-associated adenocarcinoma of the esophagus. This finding alarmingly implies that BE is biologically closer to cancer than to normal esophagus, and that the cancer risk of BE is perhaps higher than we had imagined. These findings suggest that changes modulated at the molecular biologic level supervene earlier than histologic changes, and that BE is an early intermediate stage in the process of EAC.
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- 2006
27. Polo-like kinase and survivin are esophageal tumor-specific promoters
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Yulan Cheng, Tetsuo Ito, Stephen J. Meltzer, Agnes T. Berki, Yuriko Mori, Zhe Jin, Jing Yin, Fumiaki Sato, Yutaka Shimada, John M. Abraham, Bogdan C. Paun, Takatsugu Kan, James P. Hamilton, and Suna Wang
- Subjects
alpha Karyopherins ,Esophageal Neoplasms ,Karyopherin alpha 2 ,Survivin ,Biophysics ,Cell Cycle Proteins ,Polo-like kinase ,Gene delivery ,Biology ,Protein Serine-Threonine Kinases ,Biochemistry ,Inhibitor of Apoptosis Proteins ,Proto-Oncogene Proteins ,Biomarkers, Tumor ,Humans ,RNA, Messenger ,Promoter Regions, Genetic ,Molecular Biology ,Oligonucleotide Array Sequence Analysis ,Microarray analysis techniques ,Reverse Transcriptase Polymerase Chain Reaction ,Gene Expression Profiling ,Nuclear Proteins ,Alpha Karyopherins ,Promoter ,Cell Biology ,Molecular biology ,Neoplasm Proteins ,Gene expression profiling ,Securin ,Organ Specificity ,Cancer research ,Microtubule-Associated Proteins - Abstract
For developing successful cancer gene therapy strategies, tumor-specific gene delivery is essential. In this study, we used esophageal cancer (EC) cells to identify and evaluate esophageal tumor-specific gene promoters. Four genes (polo-like kinase-1/PLK, survivin/BIRC5, karyopherin alpha 2/KPNA2, and pituitary tumor transforming gene protein 1/PTTG1) were identified by a microarray analysis as highly expressed in EC cell lines vs. five normal organ tissues (liver, lung, kidney, brain, and heart). By quantitative RT-PCR, the average mRNA expression levels of these four genes in 20 primary ECs were 2.7-fold (PLK), 6.1-fold (survivin), 2.6-fold (KPNA2), and 2.4-fold (PTTG1) higher than that of each gene in 24 different normal organs. By dual luciferase assay, the promoter activity of PLK and survivin in EC cell lines was 18.9-fold and 28.5-fold higher, respectively, than in normal lung and renal cells. The promoters of PLK and survivin could be useful tools for developing EC-specific gene therapy vectors.
- Published
- 2006
28. Molecular phenotype of inflammatory bowel disease-associated neoplasms with microsatellite instability
- Author
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Andreea Olaru, James P. Hamilton, John M. Abraham, Victoria Croog, Anca Sterian, Stephen J. Meltzer, Wolff Schmiegel, Agnes T. Berki, Yuriko Mori, Jing Yin, Noam Harpaz, Fumiaki Sato, Yan Xu, Takatsugu Kan, Elena Deacu, Suna Wang, and Karsten Schulmann
- Subjects
Adult ,Male ,Pathology ,medicine.medical_specialty ,Colorectal cancer ,Biology ,Frameshift mutation ,Risk Factors ,medicine ,Prevalence ,Humans ,Frameshift Mutation ,Promoter Regions, Genetic ,Adaptor Proteins, Signal Transducing ,Aged ,Aged, 80 and over ,Hepatology ,Transition (genetics) ,Gastroenterology ,Microsatellite instability ,Membrane Proteins ,Nuclear Proteins ,Methylation ,DNA Methylation ,Middle Aged ,medicine.disease ,Inflammatory Bowel Diseases ,Phenotype ,Neoplasm Proteins ,rab GTP-Binding Proteins ,DNA methylation ,DNA mismatch repair ,Female ,Carrier Proteins ,Colorectal Neoplasms ,MutL Protein Homolog 1 ,Microsatellite Repeats - Abstract
Patients with inflammatory bowel disease (IBD) are at increased risk of developing colorectal cancer (CRC). We sought to determine the frequency of high-level microsatellite instability (MSI-H) and the mutational and methylation profile of MSI-H IBD-related neoplasms (IBDNs).A total of 124 IBDNs (81 cancers, 43 dysplasias) from 78 patients were studied for the frequency of MSI-H and hypermethylation of 3 target genes: MLH1 , HPP1 , and RAB-32 . Fifteen MSI-H IBDNs were characterized according to their profile of frameshift mutations in 28 mononucleotide repeats and compared with 46 sporadic MSI-H CRCs.Nineteen of 124 IBDNs were MSI-H. The frequency of frameshift mutations in coding mononucleotide repeats was significantly lower in MSI-H IBDNs than in sporadic MSI-H CRCs for TGFBR2 (7 of 14 vs 34 of 43 samples; P = .047) and ACVR2 (3 of 14 vs 25 of 43 samples; P = .029). In contrast, ICA1 was mutated in 3 of 9 MSI-H IBDNs vs 2 of 54 sporadic MSI-H CRCs ( P = .028). HPP1 and RAB32 methylation was independent of MSI status and was observed in 4 of 59 and 0 of 64 nondysplastic mucosae, 20 of 38 and 1 of 25 dysplasias, and 28 of 61 and 20 of 60 carcinomas, respectively.The profiles of coding microsatellite mutations (instabilotypes) differ significantly between MSI-H IBDNs and MSI-H sporadic CRCs. Specifically, TGFBR2 and ACVR2 mutations are significantly rarer in MSI-H IBDNs than in MSI-H sporadic CRCs. Furthermore, HPP1 methylation occurs early, in 7% of nondysplastic and approximately half of dysplastic mucosae, whereas RAB32 methylation occurs at the transition to invasive growth, being rarer in dysplasias.
- Published
- 2005
29. A new specific gene expression in squamous cell carcinoma of the esophagus detected using representational difference analysis and cDNA microarray
- Author
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Naoki Teratani, Yutaka Shimada, Takatsugu Kan, Seiji Yamasaki, Atsushi Kawabe, Kan Kondo, Junichi Kaganoi, Stephen J. Meltzer, and Masayuki Imamura
- Subjects
Cancer Research ,Pathology ,medicine.medical_specialty ,Microarray ,Esophageal Neoplasms ,Biology ,Esophagus ,Antigens, Neoplasm ,Complementary DNA ,Cell Line, Tumor ,Gene expression ,Carcinoma ,medicine ,Biomarkers, Tumor ,Humans ,Gene ,Cells, Cultured ,Oligonucleotide Array Sequence Analysis ,Regulation of gene expression ,Reverse Transcriptase Polymerase Chain Reaction ,Membrane Proteins ,Epithelial Cells ,General Medicine ,Sequence Analysis, DNA ,medicine.disease ,Molecular biology ,Gene Expression Regulation, Neoplastic ,medicine.anatomical_structure ,Oncology ,Antigens, Surface ,Carcinoma, Squamous Cell ,Representational difference analysis ,human activities - Abstract
Objectives: To detect new specific gene expressions in squamous cell carcinoma of the esophagus. Methods: Representational difference analysis of cDNA (cDNA RDA) was applied to a human esophageal cancer cell line (KYSE170) and a human esophageal epithelial cell line (HEEC-1). Results: LAGE-1 was expressed specifically in KYSE170, but not in HEEC-1. It is also expressed in 27% of esophageal cancer cell lines (3/11) and 33% of esophageal cancer tissues (10/30), but not in other HEECs, normal esophageal epithelium, or other normal tissues except testis, ovary and kidney. The expression of LAGE-1 is strongly correlated with that of MAGE-A1 (p = 0.013, Fisher’s exact probability test). Fibronectin, cytokeratin 6B, cytokeratin 19, cyclin D2 and Ten-m2 were detected as candidates for downregulated genes. Reduced expression profiles of them were also identified using cDNA microarrays. The expression of LAGE-1 was induced by 5′-aza-2′-deoxycytidine (5Aza-dC) and trichostatin A (TSA) in esophageal cancer cell lines, which did not express LAGE-1. In HEECs, 5Aza-dC induced LAGE-1 expression, but TSA did not. Conclusions: LAGE-1 expression was detected in esophageal cancer by cDNA RDA. LAGE-1 might have the potential to be a target antigen for anti-tumoral immunotherapy in esophageal cancers because of its tumor-specific expression similar to that of MAGE-A1.
- Published
- 2005
30. Validation of intra-operative detection of paratracheal lymph node metastasis using real-time RT-PCR targeting esophageal squamous cell carcinoma
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Junichi Kaganoi, Masato Maeda, Go Watanabe, Shiro Nagatani, Masayuki Imamura, Takatsugu Kan, Yutaka Shimada, and Zhigang Li
- Subjects
Cancer Research ,Pathology ,medicine.medical_specialty ,Esophageal Neoplasms ,H&E stain ,Metastasis ,Paratracheal ,Medicine ,Humans ,Radiology, Nuclear Medicine and imaging ,Hematoxylin ,Staining and Labeling ,business.industry ,Reverse Transcriptase Polymerase Chain Reaction ,Paratracheal lymph nodes ,Micrometastasis ,Cell Cycle ,General Medicine ,medicine.disease ,Immunohistochemistry ,Staining ,Real-time polymerase chain reaction ,Oncology ,Lymphatic Metastasis ,Carcinoma, Squamous Cell ,Lymph Nodes ,business - Abstract
Background: We previously reported that there was a significant correlation between paratracheal lymph node (LN) metastasis and cervical LN metastasis in thoracic esophageal squamous cell carcinoma (ESCC) patients. The purpose of this study was to establish an intra-operative detection method of LN micrometastasis (MM) of ESCC using hematoxylin– eosin (HE) staining, immunohistochemistry (IHC) and real-time RT–PCR with a Light Cycler technique, and to evaluate which method, or combination of methods, is most suitable for intra-operative detection of paratracheal LN MM. Methods: Under informed consent, we obtained 33 dissected paratracheal LN samples from 22 operative patients with ESCC. Afterwards, one LN was separated into three parts by a sharp razor, and each part was checked for metastasis by HE staining, IHC with anti-cytokeratin antibody and real-time RT–PCR for SCC mRNA with a Light Cycler. Results: It took 3 h for detection by real-time RT–PCR, while it took 2 h by IHC. The detection rates of MM by HE staining, IHC and real-time RT–PCR were 50.0, 33.3 and 83.3%, respectively. However, there was a case of false negative detection that was not detected by IHC or PCR. Conclusion: The real-time RT–PCR method was useful for intra-operative detection of paratracheal LN metastasis. However, combination analysis of HE staining, IHC and real-time RT–PCR may be desirable because there was a case of false negative detection by IHC and real-time RT–PCR.
- Published
- 2004
31. Involvement of TSLC1 in progression of esophageal squamous cell carcinoma
- Author
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Tetsuo, Ito, Yutaka, Shimada, Yosuke, Hashimoto, Junichi, Kaganoi, Takatsugu, Kan, Go, Watanabe, Yoshinori, Murakami, and Masayuki, Imamura
- Subjects
Male ,Esophageal Neoplasms ,Tumor Suppressor Proteins ,Cell Adhesion Molecule-1 ,Immunoglobulins ,Membrane Proteins ,Mice, Nude ,Proteins ,DNA Methylation ,Transfection ,Gene Expression Regulation, Neoplastic ,Mice ,Cell Movement ,Protein Biosynthesis ,Carcinoma, Squamous Cell ,Disease Progression ,Tumor Cells, Cultured ,Animals ,Humans ,Female ,Neoplasm Invasiveness ,Gene Silencing ,Promoter Regions, Genetic ,Cell Adhesion Molecules ,Aged - Abstract
Frequent allelic losses of 11q23 in esophageal squamous cell carcinoma (ESCC) have been reported previously, but no tumor suppressor genes in this region have been identified in ESCC. TSLC1 was identified on chromosome 11q23.2 as a tumor suppressor gene in non-small cell lung cancer by functional complementation of a lung adenocarcinoma cell line. The purpose of this study is to evaluate the role of TSLC1 in ESCC. Loss of TSLC1 expression was observed by reverse transcription-PCR in 75% of the cell lines (27 of 36) and 50% of the primary tumors from ESCC patients (28 of 56). In a clinicopathological analysis, loss of TSLC1 expression correlated significantly with depth of invasion (pT) and status of metastasis (pM; P = 0.012 and 0.036, respectively). Patients with tumors lacking TSLC1 expression tended to have a poorer prognosis than those with tumors expressing TSLC1. (P = 0.079). Moreover, TSLC1 expression was an independent prognostic factor in a multivariate analysis (P = 0.049). Methylation analyses revealed that TSLC1 expression or loss correlated with the promoter methylation status, as determined by bisulfite sequencing, and that TSLC1 expression could be restored by a demethylating agent in certain cell lines. The growth of TSLC1-transfected ESCC cells was significantly suppressed both in vitro and in vivo (P0.01), possibly by a G(1) cell cycle arrest. TSLC1 expression also suppressed motility and invasion of ESCC cells in vitro significantly (P0.01). These findings suggest that loss of TSLC1 expression has an important role in tumor growth, cell motility, and invasion and is associated with aggressive tumor behavior in ESCC.
- Published
- 2003
32. Prognostic significance of dysadherin expression in esophageal squamous cell carcinoma
- Author
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Naoki Teratani, Jun Ichiro Kawamura, Masayuki Imamura, Takatsugu Kan, Michiie Sakamoto, Yosuke Hashimoto, Go Watanabe, Setsuo Hirohashi, Yutaka Shimada, Yoshinori Ino, Toshiya Soma, Tomoyuki Okumura, and Kan Kondo
- Subjects
Male ,Cancer Research ,Pathology ,medicine.medical_specialty ,Esophageal Neoplasms ,Biology ,Ion Channels ,Metastasis ,Cell membrane ,Downregulation and upregulation ,Predictive Value of Tests ,medicine ,Biomarkers, Tumor ,Humans ,Aged ,Neoplasm Staging ,Proportional Hazards Models ,chemistry.chemical_classification ,Membrane Glycoproteins ,Cell adhesion molecule ,Cadherin ,Esophageal disease ,Microfilament Proteins ,General Medicine ,Esophageal cancer ,Middle Aged ,medicine.disease ,Cadherins ,Prognosis ,Immunohistochemistry ,Survival Analysis ,Neoplasm Proteins ,Gene Expression Regulation, Neoplastic ,medicine.anatomical_structure ,Oncology ,chemistry ,Cancer research ,Carcinoma, Squamous Cell ,Female ,Glycoprotein - Abstract
Objective: Dysadherin is a cancer-associated cell membrane glycoprotein that has been reported to downregulate E-cadherin expression and promote metastasis. To evaluate the role of dysadherin in metastasis of esophageal squamous cell carcinoma (ESCC), we examined dysadherin and E-cadherin expression in patients with this cancer. Methods: Dysadherin and E-cadherin expression was evaluated in 117 ESCC patients (pT1, 31; pT2, 30; pT3, 39; pT4, 17) by immunohistochemistry. The findings were compared with the clinicopathological data of the patients. Results: Both dysadherin and E-cadherin were localized to the cell membrane. Thirty patients (29.1%) had tumors positive for dysadherin and 41 patients (35.0%) had tumors positive for E-cadherin. Tumors showing dysadherin positivity and negative E-cadherin expression had a significantly worse prognosis than other tumors. When the patients with dysadherin- positive tumors were combined with E-cadherin-negative patients, this group had a worse prognosis (p < 0.0001). Cox multivariate analysis revealed that dysadherin expression was an independent prognostic factor for ESCC (p = 0.003), but E-cadherin expression was not. Conclusion: Combined analysis of dysadherin and E-cadherin expression may help to predict the prognosis of patients with ESCC. Our results suggested that expression of dysadherin by this cancer may partly explain the poor prognosis of patients with preservation of E-cadherin expression.
- Published
- 2003
33. Cell culture in esophageal squamous cell carcinoma and the association with molecular markers
- Author
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Yutaka, Shimada, Masato, Maeda, Go, Watanabe, Seiji, Yamasaki, Izumi, Komoto, Junichi, Kaganoi, Takatsugu, Kan, Yosuke, Hashimoto, Issei, Imoto, Johji, Inazawa, and Masayuki, Imamura
- Subjects
Male ,Time Factors ,Esophageal Neoplasms ,Nuclear Proteins ,Proto-Oncogene Proteins c-mdm2 ,Middle Aged ,Prognosis ,Immunohistochemistry ,Proto-Oncogene Proteins ,Biomarkers, Tumor ,Tumor Cells, Cultured ,Humans ,Regression Analysis ,Female ,Neoplasms, Squamous Cell ,Tumor Suppressor Protein p53 ,Aged ,Proportional Hazards Models ,Retrospective Studies - Abstract
We reported previously that the patients in whom cancer cells could be cultured as continuous cell lines had a poor prognosis of esophageal squamous cell carcinoma (ESCC) patients. In this study, to evaluate additional evidence of prognostic significance and the genetic background of cell culture, we analyzed 203 ESCC patients.Culture samples were obtained from resected 203 primary ESCC (from 1986 to 1998; R0 resection). The expression of six molecular markers was evaluated retrospectively in resected primary esophageal tumors by immunohistochemical analysis, and the capability of establishing cell lines was compared.Thirty-five cell lines (17.2%) were established from 203 ESCC patients: group 1 (n = 35), from whom cancer cells could be cultured as continuous cell lines, and group 2 (n = 168), from whom cell lines could not be established. The cumulative survival rate of patients in group 1 was significantly lower than that of those in group 2 (P = 0.0006). Cox's proportional hazard model revealed that cell culture capability was an independent prognostic factor (risk ratio, 1.98; P = 0.007). Univariate logistic regression analysis revealed that cell culture capability had associations with the following molecular biological factors: cyclin D1, p53, murine double minute 2, p27, and fragile histidine triad gene (P0.05). However, multivariate logistic regression analysis revealed that p53 protein accumulation and MDM2 protein expression predict establishment of cell line in ESCC (odds ratio, 7.72 and 8.62, respectively).Cell culture capability is a significant prognostic factor in ESCC. p53 and MDM2 may have a crucial role in the establishment of ESCC cell lines.
- Published
- 2003
34. Indications for abdominal para-aortic lymph node dissection in patients with esophageal squamous cell carcinoma
- Author
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Yutaka Shimada, Masato Maeda, Zhigang Li, Shiro Nagatani, Junichi Kaganoi, Yosuke Hashimoto, Takatsugu Kan, Masayuki Imamura, and Fumiaki Sato
- Subjects
Adult ,Male ,medicine.medical_specialty ,Esophageal Neoplasms ,Metastasis ,Antigens, Neoplasm ,Carcinoma ,medicine ,Humans ,Aorta, Abdominal ,Prospective Studies ,RNA, Messenger ,Esophagus ,Lymph node ,Serpins ,Aged ,Retrospective Studies ,business.industry ,Esophageal disease ,Reverse Transcriptase Polymerase Chain Reaction ,Esophageal cancer ,Middle Aged ,medicine.disease ,Immunohistochemistry ,Surgery ,Dissection ,medicine.anatomical_structure ,Epidermoid carcinoma ,Lymphatic Metastasis ,Carcinoma, Squamous Cell ,Keratins ,Lymph Node Excision ,Female ,Radiology ,business - Abstract
Background. Abdominal para-aortic lymph node (APAL) dissection of esophageal cancer is not widely accepted. The aim of this article is to propose the indications for APAL dissection in esophageal cancer patients from the viewpoint of micrometastases. Methods. To evaluate the value of APAL dissection in patients with esophageal cancer, the status of APAL metastases and recurrence in 230 patients with esophageal squamous cell carcinoma (1989 to 1998) was examined retrospectively. On the basis of our findings, 16 patients received a prophylactic APAL dissection from January 1999 to March 2001. Micrometastases in the dissected lymph nodes were examined using cytokeratin staining and reverse transcription-polymerase chain reaction of squamous cell carcinoma antigen messenger RNA. Results. Among the 230 patients who had esophageal squamous cell carcinoma, 21 had APAL metastases (including micrometastases) or APAL recurrence. Among the 21 patients with APAL metastases and recurrence, 20 (95.2%) had metastases (including micrometastases) in perigastric lymph nodes (paracardial and lesser curvature nodes). Among 51 patients with lower thoracic esophageal carcinoma, 13 (25.5%) had APAL metastases or recurrence. On the basis of these results, prophylactic APAL dissection was performed in patients with lower thoracic esophageal cancer who were suspected of perigastric lymph node metastases during operations. APAL metastases (including micrometastases) were detected in 6 (38%) of these patients, and 2 patients with APAL micrometastases survived without recurrence. However, 7 patients had hematogenic recurrence after the operation. Conclusions. Our results suggested that the indications for APAL dissection were limited. Patients with lower thoracic esophageal cancer who are suspected to have perigastric lymph node metastasis and APAL micrometastases may be considered for APAL dissection. (Surgery 2002;132:93-9.)
- Published
- 2002
35. Gene expression profiling in human esophageal cancers using cDNA microarray
- Author
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Junichi Kaganoi, Yutaka Shimada, Atsushi Kawabe, Masayuki Imamura, Atsushi Itami, Masato Maeda, Seiji Yamasaki, Takatsugu Kan, and Fumiaki Sato
- Subjects
Microarray ,Esophageal Neoplasms ,Cell ,Biophysics ,Down-Regulation ,Receptors, Cytoplasmic and Nuclear ,Biology ,Adenocarcinoma ,Bioinformatics ,Biochemistry ,Polymerase Chain Reaction ,Complementary DNA ,Gene expression ,medicine ,Tumor Cells, Cultured ,Humans ,RNA, Neoplasm ,Molecular Biology ,Oligonucleotide Array Sequence Analysis ,Gene Expression Profiling ,Cancer ,Cell Biology ,Esophageal cancer ,medicine.disease ,Neoplasm Proteins ,Up-Regulation ,Gene expression profiling ,medicine.anatomical_structure ,Cancer research ,Carcinoma, Squamous Cell ,DNA microarray ,Transcription Factors - Abstract
Human esophageal cancer cell lines and human esophageal cancer tissues were profiled on cDNA microarrays. In esophageal cancer cell lines, KYAE and OE-33 (adenocarcinomas) were distinguished from KYSE series (squamous cell carcinomas). Although SK-GT-4 and TE7 were derived from adenocarcinomas, they had a comparatively similar expression profile to the KYSE series. A set of genes whose expression commonly either increased or decreased in cancer cell lines was identified. Genes that were characteristically expressed in KYAE and OE-33 were also identified. The gene expression profiles of cancer tissues (CTs) were remarkably different from those of the cancer cell lines (CCLs). Notable differences between CCLs and CTs were observed in matrix metalloproteinases, plasminogen activator, collagens, paxillin, and thrombospondin 2, etc., whose expression was not increased in CCLs but increased in CTs. Twenty-three genes were extracted to categorize patients according to their prognoses, and clustering analyses, using these genes, were performed successfully.
- Published
- 2001
36. Three-Tiered Risk Stratification Model to Predict Progression in Barrett's Esophagus Using Epigenetic and Clinical Features
- Author
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Stephen J. Meltzer, Alexandru Olaru, Yuriko Mori, Karsten Schulmann, Tsung Teh Wu, Zhe Jin, Harris G. Yfantis, Yulan Cheng, Jian Yang, Bruce D. Greenwald, Stefan David, Mary Fredericksen, Fumiaki Sato, Ziding Feng, Yvonne Romero, Bogdan C. Paun, Jean Wang, James P. Hamilton, Marcia I. Canto, Kenneth K. Wang, Tetsuo Ito, Takatsugu Kan, and John M. Abraham
- Subjects
Risk ,medicine.medical_specialty ,Esophageal Neoplasms ,lcsh:Medicine ,Oncology/Gastrointestinal Cancers ,Methylation ,Risk Assessment ,digestive system ,Gastroenterology ,Disease-Free Survival ,Mathematics/Algorithms ,Epigenesis, Genetic ,Barrett Esophagus ,Internal medicine ,Biopsy ,medicine ,Humans ,Epigenetics ,Esophagus ,lcsh:Science ,Veterans Affairs ,Molecular Biology/DNA Methylation ,Multidisciplinary ,medicine.diagnostic_test ,business.industry ,lcsh:R ,Reproducibility of Results ,Cell Differentiation ,Endoscopy ,medicine.disease ,digestive system diseases ,Treatment Outcome ,surgical procedures, operative ,medicine.anatomical_structure ,ROC Curve ,Dysplasia ,Barrett's esophagus ,Disease Progression ,lcsh:Q ,Gastroenterology and Hepatology/Esophagus ,business ,Risk assessment ,Precancerous Conditions ,Research Article - Abstract
Background: Barrett’s esophagus predisposes to esophageal adenocarcinoma. However, the value of endoscopic surveillance in Barrett’s esophagus has been debated because of the low incidence of esophageal adenocarcinoma in Barrett’s esophagus. Moreover, high inter-observer and sampling-dependent variation in the histologic staging of dysplasia make clinical risk assessment problematic. In this study, we developed a 3-tiered risk stratification strategy, based on systematically selected epigenetic and clinical parameters, to improve Barrett’s esophagus surveillance efficiency. Methods and Findings: We defined high-grade dysplasia as endpoint of progression, and Barrett’s esophagus progressor patients as Barrett’s esophagus patients with either no dysplasia or low-grade dysplasia who later developed high-grade dysplasia or esophageal adenocarcinoma. We analyzed 4 epigenetic and 3 clinical parameters in 118 Barrett’s esophagus tissues obtained from 35 progressor and 27 non-progressor Barrett’s esophagus patients from Baltimore Veterans Affairs Maryland Health Care Systems and Mayo Clinic. Based on 2-year and 4-year prediction models using linear discriminant analysis (area under the receiver-operator characteristic (ROC) curve: 0.8386 and 0.7910, respectively), Barrett’s esophagus specimens were stratified into high-risk (HR), intermediate-risk (IR), or low-risk (LR) groups. This 3-tiered stratification method retained both the high specificity of the 2-year model and the high sensitivity of the 4-year model. Progression-free survivals differed significantly among the 3 risk groups, with p=0.0022 (HR vs. IR) and p,0.0001 (HR or IR vs. LR). Incremental value analyses demonstrated that the number of methylated genes contributed most influentially to prediction accuracy. Conclusions: This 3-tiered risk stratification strategy has the potential to exert a profound impact on Barrett’s esophagus surveillance accuracy and efficiency.
- Published
- 2008
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