Search

Supplementary Figure 3 from Amplification and Overexpression of CTTN (EMS1) Contribute to the Metastasis of Esophageal Squamous Cell Carcinoma by Promoting Cell Migration and Anoikis Resistance

Supplementary Figure 2 from Amplification and Overexpression of CTTN (EMS1) Contribute to the Metastasis of Esophageal Squamous Cell Carcinoma by Promoting Cell Migration and Anoikis Resistance

Supplementary Figure 1 from Amplification and Overexpression of CTTN (EMS1) Contribute to the Metastasis of Esophageal Squamous Cell Carcinoma by Promoting Cell Migration and Anoikis Resistance

Supplementary Figure Legends 1-3 from Amplification and Overexpression of CTTN (EMS1) Contribute to the Metastasis of Esophageal Squamous Cell Carcinoma by Promoting Cell Migration and Anoikis Resistance

Gain of chromosome 11q13 is a common event in esophageal squamous cell carcinoma (ESCC). The cortactin gene (CTTN, also EMS1), located at 11q13, plays a pivotal role in coupling membrane dynamics to cortical actin assembly. This gene has been implicated in the motility of several types of cells. In the present study, we found that the amplification and overexpression of the CTTN gene was associated with lymph node metastasis in ESCC. Functional analysis by small interfering RNA–mediated silencing of CTTN revealed that in addition to the effect on cell migration, CTTN influenced cell invasiveness by anoikis resistance. In vivo assay showed that inhibition of CTTN expression also decreased tumor growth and lung metastasis of ESCC cells. At the molecular level, we showed for the first time that the protective role of CTTN in anoikis resistance was correlated with the activation of the phosphatidylinositol 3-kinase/Akt pathway. Overall, the data suggest that CTTN is an oncogene in the 11q13 amplicon and exerts functions on tumor metastasis in ESCC. (Cancer Res 2006; 66(24): 11690-9)

Si-PIN photodetectors having features such as low cost, small size, low weight, low voltage, and low power consumption are widely used as radiation detectors in electronic personal dosimeters (EPDs). The technical parameters of EPDs based on the Si-PIN photodetectors include photon energy response (PER), angular response, inherent error, and dose rate linearity. Among them, PER is a key parameter for evaluation of EPD measurement accuracy. At present, owing to the limitations of volume, power consumption, and EPD cost, the PER is usually corrected by a combination of single-channel counting techniques and filtering material methods. However, the above-mentioned methods have problems such as poor PER and low measurement accuracy. To solve such problems, in this study, a 1024-channel spectrometry system using a Si-PIN photodetector was developed and full-spectrum measurement in the reference radiation fields was conducted for radiation protection. The measurement results using the few-channel spectroscopy dose method showed that the PER could be controlled within ± 14% and ± 2% under the conditions of two and three energy intervals, respectively, with different channel numbers. The PER measured at 0° angle of radiation incidence meets the − 29% to +67% requirements of IEC 61526:2010. Meanwhile, the channel number and counts-to-dose conversion factors formed in the experiment can be integrated into an EPD.

In the research community, resistance to apoptosis is often considered a hallmark of cancer. However, pathologists who diagnose cancer via microscope often see the opposite. Indeed, increased apoptosis and mitosis are usually observed simultaneously in cancerous lesions. Studies have shown that increased apoptosis is associated with cancer aggressiveness and poor clinical outcome. Furthermore, overexpression of Bcl-2, an antiapoptotic protein, is linked with better survival of cancer patients. Conversely, Bax, CD95, Caspase-3, and other apoptosis-inducing proteins have been found to promote carcinogenesis. This notion of the role of apoptosis in cancer is not new; cancer cells were found to be short-lived 88 years ago. Given these observations, resistance to apoptosis should not be considered a hallmark of cancer.

This study was designed to evaluate the expression of C-erbB-2 and p16 in lung cancers using tissue microarray technology and to determine their clinical and pathological significance. Immunohistochemical C-erbB-2 and p16 expressions and their associations with clinical and pathological features were analyzed in two tissue microarrays. The membranous and cytoplasmic expression rates of C-erbB-2 were 40.5 and 66.5% in non-small cell lung cancers (NSCLCs), and 0 and 9.5% in small cell lung cancers (SCLCs), respectively. The nuclear and cytoplasmic expression rates of p16 were 11.5 and 32.2% in NSCLs, and 45 and 80% in SCLCs, respectively. The cytoplasmic expression of both C-erbB-2 and p16 was more frequent than the membranous expression of C-erbB-2 and the nuclear expression of p16. The rates of overexpression of C-erbB-2 and loss of p16 expression were significantly higher in NSCLCs than in SCLCs (P < 0.05). Neither C-erbB-2 nor p16 expression was significantly associated with age, tumor grade or stage, presence of lymph node metastasis or survival duration. The abnormal expressions of p16 and C-erbB-2 may play a role in the progression of lung cancers. The variations in the expression patterns of C-erbB-2 and p16 between NSCLCs and SCLCs may aid the molecular classification of lung cancer. The abnormal expression of p16 may be involved in the development of NSCLCs, and the overexpression of C-erbB-2 in NSCLCs indicates that it can be a candidate target for gene therapy.

To investigate an optimal examination method to detect micrometastases in sentinel lymph nodes (SLNs) of breast cancer.Firstly, the SLNs of breast cancer were found by 99mTc-DX isotope method. Secondly, all the SLNs which were negative by routine HE examination were serially sectioned at a 100 microm interval and stained by both HE and immunohistochemistry for detecting micrometastases. All tumor tissue paraffin blocks were also sectioned and stained with HE and immunohistochemistry as control.Totally, 121 SLNs and 44 tumors of 59 patients were examined. Micrometastasis was found to be positive in 17 SLNs (14.0%) of 14 patients (23.7%). When examined number of sections was increased from one to three, more positive micrometastatic SLNs were detected by HE staining only (3, 7, 10 for 1, 2, 3 sections, respectively). When HE staining was combined with immunohistochemical staining for AE1/3 or CK19 or muc1, much more positive micrometastatic SLNs were found (14, 12, 16 for 1, 2, 3 sections, respectively). The more sections were examined, the more micrometastases in SLNs were found. Furthermore, micrometastasis was also found to be positively correlated with the tumor size and the expression of c-erbB2, MMP-2, VEGF. The larger the tumor size was or the stronger expression of the above mentioned biomarkers, the more micrometastases in SLNs could be found.Serially sections at a 100 microm interval and staining with both HE and immunohistochemical technique using muc1 antibody may be the best way to detect micrometastases in sentinel lymph nodes in breast cancer patients.

To investigate the clinicopathological characteristics and prognostic factors in patients with intra-abdomen extragastrointestinal stromal tumors (EGISTs).The data of 47 patients of mesenchymal neoplasms that arose from the abdominal cavity and retroperitoneum, collected from July 1987 to June 2003 in our hospital with complete clinical and pathological data, were investigated retrospectively. EGISTs were diagnosed by reviewing the tumor slides stained with hematoxylin and eosin (HE). Immunohistochemistry staining were performed on CD117, CD34, smooth muscle actin, Desmin and S-100 proteins. The relations of various clinicopathologic characteristics and outcomes were examined.Among the 47 cases, 30 tumors were confirmed to be EGISTs. Twelve cases arose from the mesentery, six from small omentum, eight from retroperitoneum and four from the abdominal cavity. The size of tumors ranged from 4 to 30 cm (median 12.5 cm) in diameter and the tumor cell components mainly included spindle cells (23 cases), epithelioid cells (4 cases), and mixed cells (3 cases). The follow-up rate was 90% and the median follow up time was 44 months. The patient survival rates at 1, 5 and 10 years were 79.7%, 59.5% and 45.4% respectively. Univariate analysis showed that tumor size10 cm, tumor necrosis, mitosesor =5/50HPF, obvious nuclear atypia, moderate and poor differentiated tumor cells were predictors of poor prognosis.EGISTs have specific clinical behaviors. The parameters used for predicting GISTs prognosis are not completely applicable for EGISTs. Tumor necrosis, obvious nuclear atypia and mitosesor =5/50HPF help to predict aggressive behaviors in EGISTs.

We presented an unusual case with coexistence of carcinoid, signet-ring cell carcinoma (SRC) and heterotopic pancreatic tissue in stomach. Gastroscopic examination of this 63-year-old male patient showed multiple protrusions in gastric corpus near the greater curvature, identified by subsequent biopsy as carcinoid. Distal subtotal gastrectomy was performed. Histological and immunohistochemical examinations showed a carcinoid tumor in gastric corpus near the greater curvature, an intramucosal SRC at the lesser curvature of corpus and heterotopic pancreatic tissue in muscularis propria of the antrum at the lesser curvature with hyperplasia of peripheral endocrine cells producing multiple pancreatic hormones. We reviewed the literatures on clinicopathological characteristics and the differential diagnosis of the above three abnormalities, and concluded that the carcinoid in corpus near the greater curvature and SRC in the lesser curvature are independent lesions; the foci of endocrine cells in the muscularis propria and serosa are hyperplastic lesions from the heterotopic pancreatic tissue, rather than dissemination of carcinoid in corpus.

The optimal treatment for primary non-Hodgkin's lymphoma (NHL) of the nasal cavity remains controversial. This study was to analyze the initial response rate of radiotherapy and chemotherapy, and the influence of different treatment modalities on prognosis.From January 1996 to December 2002, the clinical data of 129 patients with previously untreated nasal NHL were retrospectively reviewed with all lesions confirmed by pathology. 116 patients were morphologically diagnosed as having nasal NK/T cell lymphoma. The immunophenotype was available in 57 cases and 52 (91.2%) of them were confirmed as NK/T-cell lymphoma. According to the Ann Arbor Staging System, 102 patients had stage I(E), 22 stage II(E), and 5 stage IV(E) disease. Among the 124 patients with stage I(E) and II(E) diseases, 22 patients received radiotherapy alone, 7 chemotherapy alone, and 95 combined modality therapy (CMT). Of these 95 patients treated with CMT, 45 patients were treated with radiotherapy followed by chemotherapy, and 50 with chemotherapy followed by radiotherapy. The primary treatment for stage IV(E) patients was chemotherapy with or without radiotherapy to the primary tumor.The overall 5-year survival (OS) and disease free survival (DFS) for all patients was 68.0% and 55.8%, respectively. It was 71.7% and 60.9% for stage I(E), and 70.6% and 47.0% for stage II(E), respectively (P0.05). The OS and DFS at the 5th year were 83.1% and 68.0% for patients who achieved complete response (CR), and 18.0% and 15.5% for those who did not, respectively (P = 0.000). Of the 124 patients with stage I(E) and II(E) disease, 67 patients were treated with radiotherapy alone (22 patients) or radiotherapy followed by chemotherapy (45), whereas 57 were treated with chemotherapy followed by radiotherapy (50) or chemotherapy alone (7). The CR rate after radiotherapy was 74.7%, however, it was only 19.3% after chemotherapy (P = 0.000). Of the 46 patients with PR, SD or PD after chemotherapy, 42 still had locoreginally localized lesion and 31 of these patients achieved CR by following radiotherapy which revealed satisfactory results. For stage I(E) and II(E) disease, the 5-year OS and DFS were 76.0% and 65.0% for radiotherapy with or without chemotherapy, and 74.4% and 56.2% for chemotherapy followed by radiotherapy. The difference was statistically not significant. However, 7 stage I(E) and II(E) patients were treated with chemotherapy alone, and 4 of them died of disease progression, with 1-year survival of 26.7%.The majority of Chinese patients with primary nasal NHL are NK/T cell in origin. The complete response rate by radiotherapy is much higher than that by chemotherapy. The addition of chemotherapy to radiotherapy did not improve the survival of patients with early stage nasal lymphoma. Radiotherapy is suggested as the primary treatment for stage I(E) and II(E) nasal NK/T cell lymphoma.

To determine the diagnostic value of B72.3, BerEP4 and calretinin in differentiating metastatic carcinoma cells from reactive mesothelial cells (RMC) in serous effusions by using immunocytochemical method (ICC), and to investigate the feasibility of ThinPrep (TP) preparation for ICC.One hundred fifty eight serous effusion specimens were examined by ICC on cell block (CB) sections (CB-ICC) using antibodies against of B72.3, BerEP4 and calretinin. Fourty-nine of the samples, ICC on ThinPrep slides (TP-ICC) and CB-ICC were performed concurrently.The sensitivities of B72.3 and Ber-EP4 for detecting carcimoma cells were 76.9% and 69.2% respectively, and when combined the sensitivity was increased to 89.7%. The sensitivity and specificity of Calretinin for detecting mesothelial cells were 90.9% and 87.2% respectively. The sensitivity of B72.3 in differentiating cancer cells from reactive mesothelial cells by CB-ICC and TP-ICC was 78.9% and 68.4%. It was 78.9% and 68.4% of BerEP4 respectively. No statistical significance was observed between CB-ICC and TP-ICC in differentiating metastatic carcinoma cells from reactive mesothelial cells.The combination of antibodies of B72.3, Ber-EP4 and calretinin is quite helpful as an auxiliary in differentiating metastatic carcinoma cells from reactive mesothelial cells. ThinPrep preparation slides may effectively replace the cell block sections for ICC in differential diagnosis of serous effusions.

To explore the prognostic factors in patients with gastrointestinal stromal tumors of the small intestine.Tumor slides stained with hematoxylin and eosin from these patients were reviewed. Two histomorphologically representative areas were identified and arrayed on a tissue microarray. Immunohistochemistry staining were performed using antibodies to detect the expression of c-kit protein (CD117), CD34, smooth muscle actin, desmin, S-100, Ki-67, P53 and bcl-2 protein. The relationship between clinicopathologic features and prognosis was analyzed by univariate analysis.The 1-, 3-, 5-year survival rate of 58 such patients were 98.3%, 69.7%, and 50.9% respectively. The prognosis was related with tumor size and gender by univariate analysis (P0.05).More attention should be paid to the male patients with small intestine stromal tumors,especially those with tumors size5 cm, because those tumors are more likely to metastasize than smaller tumors (or = 5 cm).

To investigate micro-metastasis in mediastinal lymph nodes (mLN) of patients with clinical stage I approximately II lung cancer and its clinical significance.A total of 181 mLN from 32 lung cancer patients in clinical stage I approximately II were collected during operation and their frozen sections at two different levels were examined immunohistochemically (IHC) with an anti-epithelial cell monoclonal antibody Ber-Ep4. Routine HE staining was done for comparison. The results were processed by Chi-square tests in SPSS 10.0 soft ware.Fifteen of the 32 patients (46.9%) were found to have micro-metastasis in 21 of 181 mLN (11.6%) examined by immunohistochemical staining though routine histopathological examinations were negative. Of those 15 cases, micro-metastasis was detected in 9 only by IHC and in 6 both by IHC and HE stainings. The positive rate of micro-metastasis in N0, N1, and N2 stratified by routine pathology was 36.8% (7/19), 33.3% (2/6) and 85.7% (6/7), respectively (N0 vs N2, P0.05). When stratified according to clinical staging (cTNM), pathological staging (pTNM) and pathological staging on the basis of IHC (iTNM), the frequencies of N2 cases were 0, 18.8% and 46.9%, respectively (differences among the three groups: P0.01). Nine cases reported as N0(7) and N1(2) by routine histopathological examination were found to have micro-metastasis in mLN by IHC staining, therefore they were actually N2 cases.IHC staining with a monoclonal antibody specific for epithelial cells (Ber-Ep4) is more sensitive in the detection of mediastinal micro-metastais than routine HE staining. Underestimation of the extent of mLN metastasis by cTNM and/or pTNM stagings frequently exists in patients with clinically early lung cancer.

To explore the relationship between the overexpression of PKA RIalpha mRNA and cliniopathological parameters in lung cancer.RT-PCR was used to detect the expression of PKA RIalpha mRNA in 54 cases with human lung cancer and matched normal tissues.(1) The expression of PKA RIalpha mRNA was significantly higher in cancer tissue (66.7%) than in normal tissues (20.4%) (P0.01). (2) The expression was significantly correlated with TNM stage (P0.01), being increased with TNM stage. (3) The expression was significantly higher in patients with positive lymph nodes than in those with negative lymph nodes (P0.01). (4) There were no significant associations of PKA RIalpha mRNA expression with histological type, differentiation grade or size of the tumor.This study indicates that the overexpression of PKA RIalpha mRNA may play an important role in the progression, metastasis and prognosis of lung cancer.

To study the effect of human papillomavirus (HPV) 16 E6/E7 and TPA (12-O-tetradecanog-1-phorbol-13-acetate) on malignant transformation of human embryo oral tissue.Recombinant plasmid with HPV 16 E6/E7 was constructed and transfected into human embryo oral tissue. The oral tissue with HPV 16 E6/E7 gene or without the gene was inoculated into the hypophloeodal of right shoulder in scid mice, respectively. The study was conducted in four groups: the first group was the oral tissue transfected plasmid with HPV 16 E6/E7 plus TPA, which were inoculated into 8 scid mice; the second group was only oral tissue transfected with plasmid with HPV 16 E6/E7 into 6 scid mice; the third group was normal oral tissue plus TPA inoculated into 6 scid mice, and the final group was only normal oral tissue inoculated into 5 scid mice. Three days after inoculation, TPA was injected at the left shoulder of the mice once a week. Twelve weeks after inoculation, tumor was found in 7 scid mice from the first group. HPV 16 E6/E7 gene in tumor tissues was analyzed by PCR.The rate of tumor formation was 7/8 in the first group; no tumor was found in the other groups. Pathological diagnosis of the tumor was fibrohistiocytoma. HPV 16 E6/E7 gene was detected by PCR in tumor tissues.With the cooperating action of TPA, human oral tissue containing HPV 16 E6/E7 gene could cause malignant transformation in scid mice.

To analyze ThinPrep (TP) application by comparing TP slides with conventional smear (CS) slides in fine needle aspiration cytology.A total of 522 samples from the breast, metastatic cancer, lymph node, thyroid and salivary gland were used in parallel preparations of one TP slide and one CS slide. The paired slides were compared according to cell quality, overall cellularity, cell preservation, nuclear architecture and background.Cell quality of TP was superior to CS in the breast group (36.2%, 28.0%, P0.05) and metastatic cancer group (51.0%, 14.9%, P0.05), but inferior to CS in lymph node group (16.5%, 58.2%, P0.05). Cellularities of TP and CS were similar in breast groups (25.1%), while TP had greater cellularity than CS in metastatic cancer group (32.2%, 21.8%, P0.05). Cell preservation and abnormal architecture of TP were superior to CS in breast group (36.7%, 12.1%, P0.05) and metastasis cancer group (60.9%, 9.4%, P0.05). Cell quality of TP slide was inferior to CS in lymph node group (16.5%, 58.2%, P0.05) with 27 of the 46 cases showing tuberculosis. Cell quality of TP and CS slide was similar in the thyroid and salivary gland group (35.3%). Myoepithelial cells of fibroadenoma on the TP slide were decreased in number and, due to the increased papillary and flattened cells, it was easy to diagnose benign lesion.In the breast group and metastatic cancer group, cell quality of the TP slides is superior to CS, but they are similar in thyroid and salivary gland group. The difference of diagnosis criteria between TP and CS slides exists only in tuberculosis, partly the reactive hyperplasia cases.

To evaluate the expression of annexin II in human esophageal squamous cell carcinoma (ESCC) and its relation with clinicopathological data.The expression of annexin II mRNA and protein in paired cancer tissues and their adjacent quasi-normal tissues were detected by RT-PCR, immunohistochemical method and densitometric scanning. The relation between annexin II expression and the status of tumor differentiation was analyzed.The expression of annexin II was significantly lower in the tumor tissue than that in its paired normal counterpart both in mRNA and protein level (P0.05, P0.01). The protein expression of annexin II was significantly lower in moderately and poorly differentiated tumors than those in well differentiated ones (P0.05).Down-regulation of annexin II in esophageal carcinogenesis may play an important role in squamous cell differentiation.

To study the clinicopathological features of gastric neuroendocrine tumors.Twenty cases were reviewed. The specimens were formalin-fixed, paraffin-embedded and immunostained by S-P method.Among the twenty cases, one case was carcinoid, three were malignant carcinoids, six had small cell carcinomas and ten had mixed extocrine--endocrine carcinomas. Immunohistological examination of tumor cells found 80% positive for S-100, NSE (85%), CgA (50%), SY (50%), gastrin (30%), serotonin (65%), AE1/AE3 (50%), and CEA (80%).In the WHO classification, there are five histological types in endocrine tumors of gastrointestinal tract. They are carcinoid, malignant carcinoid, small cell carcinoma, mixed exocrine--endocrine carcinoma and tumor-like lesions. But some cases in our paper were so different that they could not be classified. The gastric endocrine tumors are different from intestinal endocrine tumors and in classification, treatment and prognosis.

In order to improve early detection and visualization of esophageal premalignant lesion and carcinoma, this study was designed to explore significance of the screening by iodine staining of endoscopic examination in the area of high incidence of esophageal carcinoma.Three screenings were completed through endoscopic examination for 3164 persons in high-risk population. Esophageal epithelium was stained with iodine in endoscopic examination. According to mucosal color and pathological result, esophageal carcinoma and premalignant lesion were confirmed.After staining, the normal epithelium became brown and precancerous or malignant epithelium manifested yellow with sharp margin. (1) 100% early esophageal cancer was iodine staining positive, all lesions were grade I to grade II. The incidence rates of early esophageal carcinoma and advanced esophageal carcinoma were 1.60%-4.59% and 0.29%-1.09%, respectively. Early detection rate of esophageal carcinoma was all over 75%. (2) 95.6% severe dysplasia were staining positive; 91.3% lesion were grade I and grade II in staining. The incidence rate of severe dysplasia was 4.49%-7.68%. (3) 96.6% moderate dysplasia was positive in iodine staining, 73.3% lesion were grade II and grade III. (4) 92.3% mild dysplasia was positive in iodine staining; most of lesions were grade III. (5) 0.9% esophagitis and 0.4% normal esophageal mucosal were grade I and grade II in iodine staining, most of lesion were negative or grade III.The results demonstrated that the screening with endoscopic staining examination could improve early detection of esophageal carcinoma and precancerous lesion. Iodine staining showed highly sensitivity and specificity for identifying these precancerous and early squamous cancer lesions; it may be helpful for diagnosis of early esophageal carcinoma and precancerous lesion.

To confirm the accuracy of the ThinPrep Pap Test (Cytyc Corp., Boxborough, Massachusetts, U.S.A.) on the basis of histologic diagnosis by biopsy and the detection of human papillomavirus (HPV) DNA in mass screening.A total of 1,997 women residing in Xiangyuan County, Shanxi Province, P.R.C., an area with a high incidence of cervical carcinoma, were enrolled in this study. We collected exfoliative cervical samples from all subjects into a liquid buffer (Preserv-Cyt [Cytyc]) and utilized for both cytologic screening using the ThinPrep Pap Test and HPV DNA testing. Subsequent colposcopic biopsies were taken on all subjects. All the tests were performed in an independent and blinded fashion. We compared the ThinPrep Pap test with colposcopic biopsy and HPV test.High grade squamous intraepithelial lesions (HSIL) (CIN 2/3) were identified in 74 (3.7%) of 1,993 women adequately assessed, and there were 12 cases of squamous cell carcinoma (SCC). The false negative rate of ThinPrep cytology was 3.2% for biopsy-confirmed CIN 3 and 9.3% for CIN 2. Twenty-seven (87%) of the 31 women with biopsy-confirmed CIN 3 and 12 (100%) of 12 with biopsy-confirmed SCC had a diagnosis of either HSIL or greater abnormalities on ThinPrep cytology. In addition, the HPV DNA detection rates offered a good correlation between cytology and biopsy.The ThinPrep Pap performed extremely well in this primary screening trial. We found a good correlation between ThinPrep cytology and colposcopic biopsy on detection of HSIL and SCC; cervical specimens collected in ThinPrep liquid buffer serve as a direct test for HPV as well.