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Objective: The purpose of this study was to investigate whether matrine can affect the phenotypes and functional maturation of dendritic cells (DC), and to secrete many cytokines, as well as the cytotoxic T lymophocyte (CTL) specific killing effect induced by gastric carcinoma dendritic cells vaccine in vitro. Materials and methods: The experimental groups consisted of three concentrations of matrine (0.5, 1.0, and 2.0 mg/mL). The DC maturation was detected by flow cytometry. The proliferation of T cells was detected by cell counting kit-8 (CCK-8) method. The secretions of cytokines by CTLs were detected by enzyme-linked immunosorbent assay (ELISA). The specific killing capacity of CTLs to target MKN45 gastric cancer cells was detected by lactate dehydrogenase (LDH) release assay. Results: The results showed that matrine could increase the expressions of CD86 and CD83 in a dose-dependent manner. Matrine could promote T cell proliferation (P). Moreover, matrine also significantly increased the levels of interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α) and interleukin-12p70 (IL-12p70), respectively (P). The therapeutic vaccination with DCs vaccine treated with lipopolysaccharide (LPS) plus matrine resulted in improved killing effect as compared to that observed with DCs treated with LPS alone (P). Conclusion: These findings define matrine as an immune adjuvant that enhances the DC activation and demonstrate a new pharmacological approach to improve the therapeutic effect of autogenous DC vaccines.

Background E2Fs are genes that regulate DNA synthesis and the cell cycle by encoding a family of transcription factors. Increasing experimental evidence has revealed that E2Fs play key roles in tumor progression in various types of cancer. Methods We investigated the survival, expression and transcriptional data of E2F1/2/4 in gastric cancer (GC) patients using the immunohistochemistry assay, Kaplan-Meier Plotter, cBioPortal, String, and GEPIA databases. The plasma of GC patients was analyzed using the real-time reverse transcription polymerase chain reaction (RT-PCR) assay. The correlation between E2F1/2/4 expression and clinical features was analyzed using the quartile method. As well, the correlation between E2F1/2/4 and GC immune infiltration was also investigated using the TIMER database. Database of Immune Cell Expression (DICE) was also used to analyze correlations between SOX4 and immune responses. Results RT-PCR and tissue immunohistochemistry confirmed that E2F1/2/4 was highly expressed in serum and GC tissue samples of GC patients, the expression of which was not affected by patient age and gender. Also, the survival analysis revealed that low levels of E2F1/2/4 expression were significantly associated with a longer overall survival (OS) in GC patients. E2F1/2/4 was correlated with patient prognosis and immune cell infiltration, including B cells, CD8+ T cells, CD4+ T cells, macrophages, neutrophils, and DCs in GC. Our findings indicated that E2F1/2/4 could be used as a prognostic biomarker and indicator of immune infiltration in GC. Conclusions This study revealed that E2F1/2/4 could be a promising indicator for tumor-associated immune infiltration and prognosis in GC patients.

We investigated the expression of dimethyladenosine transferase 1 homolog (DIMT1) in human gastric carcinoma (GC) tissues, pericarcinoma histologically normal tissues, and normal gastric tissues and explored its clinical significance. Immunohistochemistry staining was used to detect the expression of DIMT1, and the findings were compared with clinicopathological features of patients with GC. The result also was ascertained by Western blotting. The Kaplan-Meier method and log-rank test were used to compare the overall survival rate and time in the DIMT1 low-level and high-level expression groups. Immunohistochemical staining indicated that the expression of DIMT1 in GC tissues (65/75; 86.7%) was significantly more common (P.001) than that in pericarcinoma histologically normal tissues (14/75; 18.7%) and normal gastric tissues (2/12; 16.7%). High expression of DIMT1 correlated closely with differentiation (P=.023), invasion (P=.042), lymph node metastasis (P=.008), distant metastasis (P=.006), and TNM stage (P=.013). Western blotting showed that DIMT1 expression correlated positively with TNM stage and implied that more advanced TNM stage was accompanied by higher expression of DIMT1 (P.001). Kaplan-Meier survival analysis showed that high DIMT1 expression correlated significantly (P.001) with a poor prognosis. Our data suggest that DIMT1 is a useful molecular biomarker to predict tumor progression and prognosis in patients with GC.

// Debao Liu 1, 2, * , Yueying Zhang 3, * , Yan Li 4 and Kaixi Fan 1 1 Department of Oncology, Affiliated Hospital of Shandong Academy of Medical Sciences, Jinan, Shandong, China 2 School of Medicine and Life Sciences, University of Jinan-Shandong Academy of Medical Sciences, Jinan, Shandong, China 3 Department of Pathology and Pathophysiology, Institute of Basic Medicine, Shandong Academy of Medical Sciences, Jinan, Shandong, China 4 Department of Central Laboratory, Affiliated Hospital of Shandong Academy of Medical Sciences, Jinan, Shandong, China * These authors have contributed equally to this work Correspondence to: Kaixi Fan, email: fankaixiyx@sina.com Keywords: gastric cancer, neurofibromatosis type I, TNM stage, immunohistochemistry, prognosis Received: December 02, 2016 Accepted: July 06, 2017 Published: August 03, 2017 ABSTRACT We investigated whether the Neurofibromatosis type-1 ( NF1 ) gene was of prognostic relevance to gastric cancer (GC) patients. Immunohistochemical staining of 160 matched GC tumor and adjacent normal tissue samples showed that 58.1% (93/160) of GC samples were NF1-positive as compared to 94.4% (151/160) of normal tissue samples (χ 2 =58.05, P 0.05). Kaplan-Meier analysis revealed that negative or low NF1 were associated with poor overall survival (OS) in gastric cancer patients ( P

Synthesis of linear and angular furocoumarins and difurocoumarins has been carried out starting from substituted coumarins and phenyl acetylene leading to the target compounds via styrylcoumarin intermediates. The average yields are higher than those reported before. Newly synthesized furocoumarin and difurocoumarin derivatives were evaluated for inhibition of cell proliferation of human breast carcinoma, human gastric carcinoma, and human lung cancer, exhibiting antiproliferative activity. New difurocoumarin derivative 4-methyl-6,9-diphenyl-2H-difuro[3,2-f:2',3'-h]chromen-2-one exhibited the highest inhibition of cell proliferation on all cell lines.

In this study, three different carboxymethyl chitosans (CMCs), namely O-carboxymethyl chitosan (OC), N-carboxymethyl chitosan (NC) and N,O-carboxymethyl chitosan (NOC), were prepared as nano-carriers for an anti-tumor drug, docetaxel (DCT). Three CMCs were fabricated into drug-loading nanoparticles under different preparing conditions respectively. A series of characterization parameters were determined including drug loading capacity (DC), entrapment efficiency (EE), particle size distribution, zeta potential, and morphology, which were used to find the optimized preparing condition for three kinds of nanoparticles (NPs). Furthermore, the in vitro drug release profiles of all the three kinds of nanoparticles showed a sustained release behavior and the release rate of OC-NPs was the fastest. Moreover, compared with NC-NPs and NOC-NPs, OC-NPs exhibited more cellular accumulation in the human gastric carcinoma cells (SGC 7901) as quantitatively determined by the flow cytometry and also much stronger cytotoxicity against these cells. Accordingly, our results showed that drug-loaded nanoparticles formed by different CMCs have dissimilar physiochemical characteristics which subsequently influence their therapeutic effects. We suggest that OC-based nanoparticle is a more desirable anti-tumor drug delivery system than NC-NP and NOC-NP.

Inonotus obliquus is a medicinal mushroom with a variety of biological activities. It has reported to have strong anti-cancer, antioxidant and anti-inflammatory properties. EBV+ gastric carcinoma is one of the most common EBV-associated cancers that were caused by latent EBV infection. In this study, we investigated the anti-cancer effects of ethanol extract of I. obliquus using in vivo xenograft animal models implanted with EBV+ human gastric carcinoma (SNU719). We also explored the molecular mechanisms responsible for its anti-cancer activity. The result indicated that the extract of I. obliquus had an anti-cancer effect in in vivo xenograft mice with EBV+ gastric carcinoma (SNU719). Extract of I. obliquus also showed a great effect on inducing the expression of p53, p21 and Bax in tumor tissue derived from EBV+ human gastric carcinoma, and these were correlated with increased expressions of the cleaved forms of caspase-9 and Parp. Also, I. obliquus attenuated the expression of viral proteins, BZLF-1 and LMP-2 in tumor tissue from EBV+ human gastric carcinoma.

The aim of the study is to analyse the expression of Interleukin-6 receptor in different human gastric tissue and to correlate with the clinicopathological features of the patients. Immunohistochemistry was done against the IL-6R antibody and the Q-score was calculated from the staining pattern. Higher Q-scores were observed in tumour cells than the adjacent normal cells which were statistically significant. We also observed a significant correlation between the expressions of IL-6R and the clinicopathological features These findings suggest that IL-6R may represent as a therapeutic target for gastric carcinoma and serve as a prognostic indicator, as well.

Two series of thieno[2,3-d]pyrimidine derivatives were designed and synthesized, in which bioactive α -aminophosphonate subunits were introduced at the N3 position through an N N bond connection. The in vitro cytotoxic activity of the novel compounds was tested against human esophageal carcinoma cells (EC109), human hepatocarcinoma cells (HepG2), human gastric carcinoma cells (MGC-803), respectively, by the MTT method. The evaluation results revealed that compounds 6mb , 6mf , 6mg , 6nd and 6nh exerted the most potent inhibition against HepG2, MGC-803 and EC109 cells, respectively. In particular, compound 6mg presented excellent inhibitory effect against HepG2 (91.2%) and MGC-803 (94.4%) cells.

Medical applications of nonequilibrium atmospheric pressure plasma (NEAPP) have been reported in cancer therapy. Cells with tumorigenic potential are limited to a small population, called cancer-initiating cells (CICs), which are usually expressing aldehyde dehydro- genase (ALDH) in a high level. CICs are believed to cause recurrence or metastasis due to their resistance to apoptosis. Here, we examined the effect of NEAPP on CICs using human uterine en- dometrioid adenocarcinoma cells and poorly differentiated human gastric carcinoma cells. When treated with NEAPP, ALDH-high cells fell into apoptosis in a comparable level to ALDH-low cells. These results suggested that NEAPP treatment was effective not only on non-CICs but also on CICs. NEAPP might become a new therapeutic approach to CICs.

Using dehydroepiandrosterone as the starting material, we have synthesized a series of steroid analogs possessing a D-ring fused with heterocycles which are pyridine, imidazo [2,1-b]thiazoles or substituted thiazole imines. All the final structures are first reported and identified by NMR and MS spectroscopys, the yields of these products are moderate to good and the reaction conditions are mild. The cytotoxicity of the synthesized compounds against EC-109(human esophageal carcinoma), EC-9706(human esophageal carcinoma), MGC-803(human gastric carcinoma) were investigated.

Licorice extracts have been widely used in herbal and folk medications. Glycyrrhiza contains diverse range of biological compounds including triterpenes (glycyrrhizin, glycyrrhizic acid) and flavonoids (quercetin, liquiritin, liquiritigenin, glabridin, licoricidin, isoliquiritigenin). The flavonoids in licorice are known to have strong anti-cancer activities. Quercetin, the most abundant flavonoid, has been shown to have anti-ulcer, anti-cancer, antioxidant, and anti-inflammatory properties. Latent Epstein-Barr virus (EBV) infection can lead to serious malignancies, such as, Burkitt’s lymphoma, Hodgkin’s disease and gastric carcinoma(GC), and (Epstein-Barr virus associated gastric carcinoma) EBVaGC is one of the most common EBV-associated cancers. In this study, the authors first examined the anti-cancer effects of quercetin and isoliquiritigenin in vivo xenograft animal models implanted with EBV(+) human gastric carcinoma (SNU719) or EBV(−) human gastric carcinoma (MKN74), and then explored the molecular mechanisms responsible for their anti-cancer activities. The results obtained showed that anti-cancer effect of quercetin was greater than isoliquiritigenin in mice injected with EBV(+) human gastric carcinoma (SNU719) cells. On the other hand, quercetin and isoliquiritigenin had similar anti-cancer effects in mice injected with EBV(−) human gastric carcinoma (MKN74) cells. Interestingly, quercetin inhibited EBV viral protein expressions, including EBNA-1 and LMP-2 proteins in tumor tissues from mice injected with EBV(+) human gastric carcinoma. Quercetin more effectively induced p53-dependent apoptosis than isoliquiritigenin in EBV(+) human gastric carcinoma, and this induction was correlated with increased expressions of the cleaved forms of caspase-3, -9, and Parp. In EBV(−)human gastric carcinoma (MKN74), both quercetin and isoliquiritigenin induced the expressions of p53, Bax, and Puma and the cleaved forms of caspase-3 and -9 and Parp at similar levels.

In most populations, gastric cancer (GC) is approximately two times more prevalent in men than in women, which may suggest the protective role of sex steroid hormones in gastric carcinogenesis. Steroid hormones such as androgens and estrogens can be synthesized not only in the gonads but also in peripheral tissues. Many researchers have conducted studies examining the expression profile of enzymes involved in steroid synthesis, the occurrence of estrogen receptors (ERs) and the influence of ERs in the development, proliferation and progression of gastric cancer. Some studies have also evaluated the relationship between the presence of ERs and survival prognosis. However, the results of these studies are often controversial and divergent. In a recent study, it was indicated that sex steroid hormones and estrogen receptors are partly involved in gastric cancer but their clinicopathological significance still needs further investigation.

Contents of phenolics and flavonoids, antioxidant activity and cytotoxicity were investigated in the methanol extracts of three different Taraxacum species, Taraxacum coreanum, Taraxacum mongolicum, and Taraxacum officinale. Total phenolics content at 1000 mg kg -1 was more present in shoot parts than in roots, and was highest in T. mongolicum shoot and root extracts (76.8 and 40.0 mg kg -1 , respectively), followed by T. coreanum and T. officinale (p -1 than total phenolics. The antioxidant activity of the methanol extracts from all the species dose-dependently increased. DPPH free radical scavenging activity at 1,000 mg kg -1 was highest in shoot and root extracts from T. mongolicum by 89.6 and 83.4%, respectively. According to MTT assay, cell viability of Calu-6 (human pulmonary carcinoma) was lowest in the T. mongolicum shoot and root extracts (IC 50 values=83.4 and 66.4 mg kg -1 ，respectively), and followed by T. coreanum and T. officinale (lowest). Calu-6 was more sensitive to the extracts than SNU-601 (human gastric carcinoma). Antioxidative and anticancer activities in three different Taraxacum species was more correlated with total phenolics content (r 2 =0.0097 to 0.6213) than with total flavonoids level (r 2 =0.0027 to 0.4627). The results showed total phenolics content and total flavonoids level were nighly correlated with anticancer activity and antioxidant activity, and their content and activities were different depending on species.

Regular use of aspirin (ASA) could reduce the risk of gastric cancer although the precise mechanism remains unclear. Down-regulation of survivin may be one of the cyclooxygenase-independent mechanisms whereby ASA induces apoptosis of gastric cancer cell. In this study, we investigated the effect of ASA on the growth, apoptosis and survivin expression of gastric cancer cell line SGC7901. The survival of cells treated with 3.0 and 10.0 mmol/L ASA for 24 h was decreased by 44.6% and 88.5%, respectively. ASA at 3.0 mmol/L inhibited the viability of SGC7901 cells in a time-dependent manner. Apoptosis analysis showed similar results with MTT assay. ASA at 3.0 and 10.0 mmol/L decreased the mRNA transcript levels of survivin and reduced survivin protein levels in SGC7901 cells also in a time-dependent manner. Our findings indicated that ASA inhibited the proliferation of SGC7901 by suppressing survivin at both the transcriptional and translational level.

Gastric cancer is a worldwide cancer with poor prognosis. Identification of diagnostic biomarkers and effective therapeutic targets is important in the treatment and diagnosis of gastric cancer. Recently, researchers have found that microRNAs play several important roles in carcinogenesis. The purpose of this study was to investigate the relationships between miR-421 expression patterns in human gastric cancer tissues with clinicopathological features.Sixty gastric carcinoma and 18 non-tumor tissues were collected from the Secondary Hospital of Ningbo, China. For quantitative detection of the expression level of miR-421, total RNA was extracted and then reverse transcription-polymerase chain reaction was performed. The relationship between miR-421 expression in gastric cancer and clinicopathological features was analyzed. After miR-421 inhibitor was transfected into gastric cancer cells, cell growth was measured by MTT assay. Finally, the expression of its target genes was detected by Western blotting.The miR-421 was over-expressed in 73.33% (44/60) of the gastric cancer samples examined. Over-expression of miR-421 in gastric cancer tissues was not found associated with clinicopathological features. The positive detection rate of miR-421 was higher than that of serum carcino-embryonic antigen (chi(2) = 39.811, P0.001). Inhibition of miR-421 expression decreased the growth of both MGC-803 and SGC-7901 gastric cancer cells in vitro, with up-regulating the expression of its cancer-related target genes, CBX7 and RBMXL1.miR-421 may involve in the early stage of stomach carcinogenesis and could be used as an efficient diagnostic biomarker.

A series of zinc phthalocyanines tetra-α-substituted with 4-(butoxycarbonyl) phenoxy groups ( 1a ) or 4-carboxylphenoxy groups ( 2a ) or 4-(2-carboxyl-ethyl)phenoxy groups ( 3a ), and the corresponding tetra-β-substituted ( 1 – 3b ) analogues, have been synthesized and characterized. The effects of the position of substituents at the phthalocyanine skeleton on their spectroscopic, photochemical and photobiological properties have been revealed. When compared with the tetra-β-substituted phthalocyanines, the corresponding tetra-α-substituted analogues exhibit a less aggregating trend in the cellular growth medium, a slightly higher singlet oxygen quantum yield and higher photo-stability in DMF, and a comparable cellular uptake. As a result, the tetra-α-substituted zinc phthalocyanines exhibit a higher photocytotoxicity toward MGC803 human gastric carcinoma cells than the tetra-β-substituted counterparts. Among all these compounds, phthalocyanine 2a shows the highest photodynamic activity, which may mainly be due to its non-aggregated nature in cellular culture medium and high cellular uptake.

Background/Aims: RUNX3 (PEBP2αC/CBFA3/AML2) is a novel tumor suppressor gene in the human gastric carcinoma. The aims of this study were to determine the methylation of RUNX3 promoter and the association between RUNX3 methylation and the clinicopathological characteristics of patients with gastric carcinoma. Methods: Seventy-nine patients with gastric carcinoma were studied prospectively from April 2005 to May 2007. The methylations of RUNX3 promoter on the gastric carcinoma specimens and the corresponding nonneoplastic mucosa were evaluated by methylation-specific polymerase chain reaction. Results: Comparison of the results with the clinicopathological characteristics identified RUNX3 monoallelic methylation in 32.9% (26/79) of the gastric carcinoma patients and in 11.4% (9/79) of those with nonneoplastic mucosa (p=0.053). The monoallelic methylated gastric carcinoma specimens predominantly consisted of well- and moderately differentiated carcinomas (44.7%), with the unmethylated group constituting 22.0% of them (p=0.031). Among the 48 patients (60.8%) who underwent gastrectomy, there was no correlation between the two groups with regard to Lauren's classification (p=0.235), depth of invasion (p=0.990), nodal status (p=0.601), stage (p=0.900), lymphatic invasion (p=0.537), and vascular invasion (p=0.815). Conclusions: Methylation of the tumor suppressor gene RUNX3 might be one of the mechanisms involved in the pathogenesis of gastric carcinoma. (Gut and Liver 2008;2:119-125)

Objective To investigate the expression of maspin, uPA and MMP-7 in gastric carcinoma and discuss their roles in tumorigenesis and progression of gastric cancer.

OBJECTIVE To explore the method of preparation of 99mTc labeled Anti-VEGF McAb 5-FU loaded polylactic acid nanoparticles (99mTc-5-FU-Ab-NPs), and investigate the biological distribution of the nanoparticles in human gas -tric carcinoma xenografts. METHODS Anti-VEGF monoclonal antibodyes (MCAB) in 5-FU-Ab-NPs were labeled with 99mTc using a modified Schwarz method. After isolation of the 99m Tc-5-FU-Ab-NPs using a Sephadex G-250 column, the labeling per-centage and radiochemical purity were determined using paper chromatog -raphy. The immunocompetence of the 99mTc-5-FU-Ab-NPs as tumor markers was determined using ELISA and immunohistochemistry. 99mTc-5-FU-Ab-NP (experimental group), 99mTc-labelled murine multiclonal IgG loaded polylactic acid and nanoparticles (control group) were injected via the tail vein into SCID mice bearing human gastric carcinoma. A radio-immunity ECT image was developed at 2 and 6 h after the injection. Following the ECT imaging, the mice were sacrificed, their tissue and tumor radioactivity distribution de termined, and percentage of the injected-dose per gram (%ID/g) and tumor/nontumor (T/NT) ratio calculated. High performance liquid chromatography (HPLC) was used to determine the 5-FU concentration in the tumor tissue and blood in the mice of both groups. RESULTS The percentage of 99mTc-5-FU-Ab-NPs labeling was 90%~95%. There was no obvious decrease in the antibody activity before and after labeling. The radio immuno-imaging (RII) showed that the tumor image had developed 2 h after injection of the 99m Tc-5-FU-Ab-NPs, and with time it was clearer at the 6th hour following the injection. The %ID/g of the tumor tissue at both 2 h and 6 h after the injection was significantly higher compared to the control group. The tumor %ID/g and the tumor to blood activity ratio (TB) of the experimental group at 6 h following the injection in -creased compared to that at 2 h, and at the same time, 5-FU concentration in the tumor of the experimental group continuously increased over time, and showed a significant difference compared to the 5-FU concentration in the tumor of the control group. CONCLUSION The 99mTc-5-FU-Ab-NPs prepared in this study are ade-quate to meet the demands of the RII, and the immune targeting ability of the anti-VEGF MCAB is reliable. Six hours after injection, the 99mTc-5-FU-Ab-NPs showed a relatively high specific concentration shadow in the human gastric carcinoma xenografts.

Osteopontin (OPN) is a secreted, integrin-binding glycophosphoprotein that has been implicated in the progression of various solid tumors. To evaluate the clinical significance of OPN in gastric carcinoma, we investigated OPN expression in resected tumors.Expression of OPN protein by gastric cancer cells was evaluated using western blot analysis. OPN messenger RNA (mRNA) expression in 18 gastric cancers was compared with that in the corresponding normal gastric epithelium by semiquantitative reverse transcription polymerase chain reaction (RT-PCR). Paraffin sections of tumors from 295 patients with gastric cancer were also investigated using immunohistochemistry.All four gastric cancer cell lines analyzed using western blotting had almost the same level of OPN protein expression as the positive control (HeLa cells). OPN mRNA was upregulated in 83% (15/18) of the tumors studied. On immunohistochemical staining, 90 tumors were classified as negative (-), whereas 205 were classified as positive (1+, 2+, or 3+). The level of OPN protein expression was significantly associated with the patient's age (p = 0.04), tumor depth (p = 0.03), histological grade (p = 0.008), and hematogenous metastasis (p = 0.007). Kaplan-Meier analysis showed that OPN positivity was significantly associated with a shorter survival time (p = 0.027). Furthermore, multivariate analysis revealed that OPN positivity was an independent risk factor for hematogenous metastasis (p = 0.034).The present findings suggest that increased tumor expression of OPN is an important determinant of shorter survival time and that OPN positivity may be useful for predicting the risk of hematogenous metastasis in gastric cancer patients.

Background and objective We developed a new imaging system to detect sentinel nodes (SNs) using a novel fluorescent tracer, ATX-S10Na(II), and investigated its usefulness in an animal model. Study design/materials and methods Human gastric carcinoma cells were implanted orthotopically into nude rats. ATX-S10Na(II) was injected subserosally into the primary tumor lesion, and visualized by a fluorescence spectro-laparoscope. Presence of tumor cells in lymph nodes (LNs) was determined by RT-PCR specific for human beta-actin. Results Injection of ATX-S10Na(II) was successful in 27 tumor-bearing rats. A red fluorescence was incorporated into the left gastric and hepatic LNs in 25 and 2 rats, respectively. Of note, human beta-actin was detected in most of these LNs. Fluorescence was not detected in LNs that did not contain cancer. Conclusion ATX-S10Na(II) is useful for the detection of cancer-containing SNs in an animal model of gastric carcinoma, and may serve as a novel tracer in SN navigation surgery.

Therapy guided by chemotherapy sensitivity and resistance assays may lead to rational treatment decisions. Paclitaxel, one of several new drugs for gastric carcinoma, has not been extensively evaluated by in vitro chemosensitivity tests.Chemosensitivity testing by histoculture drug response assay (HDRA) was performed with fresh specimens of primary tumor from 113 patients with gastric carcinoma. The test was performed in medium containing paclitaxel at three different concentrations for the initial 45 samples. Correlations between the results of chemosensitivity testing, determined at the optimal concentration, and the patients' clinicopathologic factors and outcome were then assessed for all patients.By analyzing the initial 45 samples, 10 microg/ml was considered the optimal concentration of paclitaxel for this test. HDRA was successfully performed for 100 of 113 samples and chemosensitivity, calculated as the percentage of optical density of a tumor treated with anticancer drugs in relation to the optical density of the tumor cultured in the medium only, was distributed widely at this concentration. No significant correlation was observed between chemosensitivity and age, sex, clinical stage, histopathologic type, and outcome of patients with gastric carcinoma.A histoculture drug response assay can now be performed to predict the chemosensitivity of paclitaxel at the concentration found in the current study. The accuracy of the assay to actually predict survival needs to be validated by a prospective clinical trial involving patients who have received paclitaxel in the postoperative adjuvant setting.

Therapy resistance is the major obstacle to advances in successful cancer treatment. To characterize chromosomal alterations associated with different types of acquired MDR and thermoresistance, we applied CGH to compare a unique panel of human gastric carcinoma cells consisting of the parental, drug-sensitive and thermosensitive cancer cell line EPG85-257P, the atypical MDR variant EPG85-257RNOV, the classical MDR subline EPG85-257RDB and their thermoresistant counterparts EPG85-257P-TR, EPG85-257RNOV-TR and EPG85-257RDB-TR. CGH with genomic DNA prepared from these cell lines as probes successfully identified genomic gains and/or losses in chromosomal regions encoding putative genes associated with drug resistance and/or thermoresistance. These genes included various members of the families of ABC transporters and molecular chaperones. The importance of these cell variant-specific genomic imbalances in the development of MDR and thermoresistance is discussed and remains to be elucidated. © 2002 Wiley-Liss, Inc.

Antigenic peptides have been used as a cancer vaccine in melanoma patients and have led to a drastic regression of metastatic tumors. However, few antigens have been identified in non-melanoma tumors. We recently purified a new natural antigenic peptide, designated F4. 2, by biochemical elution from a human gastric signet cell carcinoma cell line and showed that it is recognized by an autologous human histocompatibility antigen (HLA)-A31-restricted cytotoxic T lymphocyte (CTL) clone. Here we describe in vitro induction of F4. 2-specific CTLs from peripheral blood T lymphocytes of HLA-A31( +) gastric cancer patients. The T cells of seven HLA-A31( +) patients with gastric cancers were stimulated in vitro by F4.2-pulsed autologous dendritic cells which had been induced from peripheral blood of each patient by incubation in the presence of granulocyte macrophage colony-stimulating factor (GM-CSF) and IL-4. We tested the cytotoxicity of the T cells against F4.2-loaded C1R-A*31012 by a 6-h (51)Cr release assay after 3 stimulations with F4.2-pulsed dendritic cells. F4.2-specific cytotoxicity was detectable in the stimulated T cells from two of the seven HLA-A31( +) patients. Further, both F4.2-specific CTLs also lysed the gastric cancer cell line, HST-2, from which F4.2 was derived. These results suggest that F4.2 peptide may be useful as an HLA-A31-restricted peptide vaccine in certain patients with gastric cancer.

Background. Recent studies suggest that lectin binding activity is correlated with the metastasis and prognosis of several human carcinomas. Wheat germ agglutinin (WGA) is a lectin that recognizes mainly N-acetyl-glucosamin (GalNAc) and acetyl-neuramic acid. However, little is known about WGA expression in gastric carcinoma. The purpose of this investigation was to clarify the significance of WGA expression in human gastric carcinoma.

Recent epidemiological studies indicate that there is reduced risk of all digestive carcinomas in patients who regularly take nonsteroidal anti-inflammatory drugs (NSAIDs) such as aspirin. Cyclooxygenase (COX) is a target enzyme for NSAIDs. We investigated the role of two isoforms, COX-1 and COX-2, in the development and metastasis of gastric carcinoma.Fifteen gastric carcinoma tissue specimens and accompanying adjacent mucosa specimens were obtained from surgical resections. COX-1 and COX-2 protein expression were evaluated using Western blotting analysis, and their relative band densities were semi quantified using standard densitometry scanning techniques.Compared with paired noncancerous specimens, COX-2 was overexpressed in 10 of 15 carcinoma tissue specimens (66.7%). Overall, COX-2 levels in carcinoma tissue were significantly higher. Two early carcinomas (confined to the mucosa and submucosa) and three of 13 advanced carcinomas (extended below the submucosa into the muscular wall) had weak or similar COX-2 expression in paired tissue specimens. COX-2 overexpression in tumors significantly correlated with tumor invasion into the lymphatic vessels in the gastric wall and metastasis to the lymph nodes. Furthermore, the stage grouping in the TNM classification significantly correlated with COX-2 overexpression. In contrast, COX-2 overexpression did not correlate with histopathological grading, surface size, and venous vessel invasion of the tumors. COX-1 levels were similar between paired tissues.COX-2 overexpression might enhance lymphatic invasion and metastasis in patients with gastric carcinoma, implicating a poor prognosis. Therefore, the use of COX-2-specific inhibitor to suppress lymphatic metastasis in humans should be investigated.

A previous study reported a relationship between the expression of heat-shock protein 60 (HSP60) by Helicobacter pylori and its adhesion to human gastric carcinoma (MKN45) cells. To examine whether the HSP60 homologue of H. pylori is associated with the adhesion of H. pylori to human gastric epithelial cells, an inhibition assay of adhesion of H. pylori to MKN45 cells was performed by flow cytometric analysis with monoclonal antibody (MAb) designated as H20 recognising HSP60 of H. pylori. The rate of adhesion of H. pylori pretreated with MAbH20 to MKN45 cells was lower than that of untreated H. pylori. Primary human gastric epithelial cells from a patient with gastric cancer were also prepared for comparison in the inhibition assay with MAbH20. H. pylori adhered to the primary human gastric epithelial cells, and this adhesion was significantly inhibited by MAbH20. These results suggest that the H. pylori HSP60 homologue recognised by MAbH20 might be associated with the adhesion of H. pylori to primary human gastric epithelial cells as well as to cultured gastric cancer cells.

CDC25 phosphatases activate cyclin‐dependent kinases by removing inhibitory phosphate groups on the molecules and positively regulate the cell cycle progression. The expression of CDC25A, B and C was examined in gastric carcinoma cell lines and gastric carcinoma tissues by northern blotting and immunohistochemistry. The gastric carcinoma cell lines expressed CDC25A, B and C mRNA at various levels. The expression levels of CDC25B were generally higher than those of CDC25A and C. Of the 40 gastric carcinomas, 70% of the tumors expressed CDC25B mRNA at higher levels than the corresponding normal mucosas, while 38% overexpressed CDC25A mRNA. The CDC25C expression was at very low or undetectable levels. No obvious correlation was detected between the expression of CDC25B and p53 gene mutations. Inununohistochemically, CDC25‐positive tumor cells were detected in 43 (78%) of 55 gastric carcinoma cases, of which 27 (49%) were strongly positive. Strong expression of CDC25B protein was associated with advanced stage and deep invasion. Furthermore, the incidence of strong expression was significantly higher in carcinomas with nodal metastasis than in those without metastasis. These findings suggest that Overexpression of CDC25B may favor development and progression and may be an indicator of malignant behavior of gastric carcinomas.

The anti‐tumor and anti‐metastatic effects of TNP‐470, an angiogenesis inhibitor, and mitomycin C (MMC), a representative anti‐neoplastic agent, were investigated using our established liver‐metastasizing gastric carcinoma line, AZ‐H5c. AZ‐H5c was injected into the spleen of nude mice which had been randomly divided into 4 groups; a control group given saline solution, a group receiving 15 mg/kg TNP‐470, a group receiving 30 mg/kg TNP‐470 and a group receiving 2 mg/kg MMC. TNP‐470 was given s.c. on alternate days for 5 weeks from day 10 after intrasplenic injection, and MMC was administered intraperitoneally (i.p.) once a week from day 10 after intrasplenic injection. In the control group, liver metastasis developed in 13 of 16 mice (81%). Liver metastasis developed in 6 of 11 mice (55%) receiving MMC. In contrast, liver metastasis developed in 4 of 8 mice (50%) receiving 15 mg/Kg TNP‐470, and in 0 of 14 mice (0%) receiving 30 mg/kg TNP‐470. However, TNP‐470 had no effect on the tumor growth. These results indicate that the angiogenesis inhibitor TNP‐470 has a strong inhibitory activity against in vivo liver metastasis of human gastric carcinoma.

Characteristics of the synthesis and Mg-dependent phosphorylation of gastroproteins are studied in intact cells of the mucous coat of the stomach and in the cells of tumor nodes in the same patients. Comparison of polypeptide maps reveals 4 main types of mucous coats which contain p18 (phosphorylated or not), p20, or p22, or no characteristic feature at all. Two kinds of changes, namely, accessory proteins and proteins disappearing from the spectrum, are manifested mainly in extracts of primary tumors as compared to homologous mucous coats of the stomach. Three classes of gastric tumors are distinguished according to the difference between tumor and normal cell polypeptides: I) no difference, II) isolated differences, and III) numerous qualitative and quantitative differences in gastropolypepides and their phosphoforms.

There is a need to establish animal models which are suitable for investigation of human gastric cancer metastasis to the liver. To this end, a human gastric carcinoma line, AZ521 was injected into the spleens of nude mice. Cells from the few liver metastatic foci of injected AZ521 were expanded "in vitro" and subsequently injected into the spleens of nude mice. By repeating these procedures three times, we were able to obtain a cell line, designated as AZ-H3c, with high metastatic potential in nude mice. Liver metastasis developed in 15 of 21 (71%) animals injected with AZ-H3c, but only in 14% of those injected with parental AZ521. Further, AZ-H3c caused faster tumor development than did AZ521. However, the primary AZ-H3c tumors and liver metastatic AZ-H3c tumors showed essentially the same histological appearance. We also analyzed the cell surface expression of adhesion molecules. The data showed that the expression of VLA-1, VLA-2, VLA-3, VLA-4, VLA-5 was enhanced in AZ-H3c. In contrast, the expression of VLA-6, (alpha(v)beta3), E-cadherin, ICAM-1 and LFA-1 was reduced in this high-metastatic line. These results suggest that (beta1) integrins play an important role in the liver metastasis of human gastric carcinoma cells. Our high-metastatic line should be useful for studies aimed at the prevention of liver metastasis.

Compound (I) inhibits the HGF-induced scattering of MDCK (epithelial cells) and in vitro tumorigenesis of H1437 (non-small-cell lung cancer) and GTL-16 (human gastric carcinoma).

CD40 signaling plays a critical role in the survival rate of gastric cancer patients. Tumour samples were collected from 73 patients with who were diagnosed as gastric cancer in general surgery department in the 1st affiliated hospital of Suzhou University between September 2002 and July 2003. All patients had not received radiotherapy and chemotherapy before operation. These patients include 46 male and 27 female. Here we show that CD40 is constitutively expressed in the human gastric carcinoma tissues, and CD40 protein and mRNA positive expression in gastric cancer tissues closely correlated with lymph node metastasis and tumour TNM stage. CD40 positive expression in gastric cancer patients with lymph node metastasis was markedly higher than that in gastric cancer patients without lymph node metastasis. CD40 positive expression in stage III–IV gastric cancer patients was markedly higher than that in stage I–II gastric cancer patients. Moreover, CD40 expression closely correlated with prognosis of gastric cancer patients. Therefore, CD40 was taken as grouping variable, and lymph node metastasis and clinical staging were taken as stratification variables, respectively, further analysis showed that prognosis in gastric cancer patients with lymph node metastasis and CD40 positive expression was markedly worse than that in gastric cancer patients without lymph node metastasis and CD40 negative expression (P = 0.0076). These results suggest that CD40 signaling plays a critical role in the survival of gastric cancer patients.

Background The aim of this study was to clarify the clinical significance of TM4SF members CD9, CD63 and CD82 in human gastric carcinoma. Methods By employing RT-PCR and immunohistochemistry, we studied the expression of CD9, CD63 and CD82 in 49 paired tissue specimens of normal gastric mucosa and carcinoma. All tissues were obtained from patients who underwent curative surgery. Results All normal gastric epithelium and gastric ulcer tissues strongly expressed transcripts and proteins of CD9, CD63 and CD82 as compared with corresponding controls. We found a significant correlation between CD63 mRNA level and different pM statuses (P = 0.036). Carcinomas in M0 stage revealed a stronger expression of CD63 than carcinomas in M1 stage. Expression of CD9 protein was found significantly stronger in pN0, pM0 than in advanced pN stages (P = 0.03), pM1 (P = 0.013), respectively. We found the relationship between CD63 expression, gender (p = 0.09) and nodal status (p = 0.028), respectively. Additionally, advanced and metastasized tumor tissues revealed significantly down-regulated CD82 protein expression (p = 0.033 and p = 0, respectively), which correlated with the tumor pTNM stage (p = 0.001). Conclusion The reduction of CD9, CD63 and CD82 expression are indicators for the metastatic potential of gastric carcinoma cells. Unlike their expression in other tumor types, the constitutive expression of CD63 may indicate that this factor does play a direct role in human gastric carcinogenesis.

Cytotoxic activity of aqueous extracts from adult moth abdomens of 12 species belonging to the subfamily Arctiinae was tested in vitro using human gastric carcinoma TMK-1 cells. The strong cytotoxic activity, down to a dilution of 1/105, was found in females of all of 12 species of moths. However, samples of the males of the same species were without cytotoxicity. Females of three species, not of this subfamily, and those of 9 species, belonging to other family than Arctiinae, did not show any cytotoxicity. The strong cytotoxic activity in female abdomens of Hyphantria cunea was located in the ovaries. The active principle in the extract sample from H. cunea was inactivated by heat, acid or alkaline treatment and digested by proteases, indicating probably protein in its nature. Thus, cytotoxic factors in moths of the subfamily Arctiinae were likely to be derived from their ovaries.

AIM: To study the expression of survivin, an inhibitor of apoptosis protein, in human gastric carcinomas and gastric carcinoma models of rats.

A poorly differentiated medullary carcinoma of human stomach, designated HY-1, was successfully transplanted to nude mice by either the subcutaneous or intramuscular route for five generations. The transplanted tumor showed spontaneous lung metastases in nearly 100% of KSN and Balb/c female nude mice. There were over 20 visible lung metastatic nodules in KSN and Balb/c nude mice bearing tumors for over 80 days. Immunostaining of type IV collagen and electron microscopy revealed that tumor cells were often in direct contact with basement membrane (BM) of tumor blood vessels in the primary tumor tissue. At the site of contact between tumor cells and vascular BM, focal disappearance of the BM, disruption of endothelial cells and entry of tumor cell clusters into vascular lumen were observed. Immunostaining of 72 kDa gelatinase/type IV collagenase demonstrated that tumor cells expressed this enzyme in their cytoplasm. These results suggest that spontaneous metastasis of this tumor may be partly due to a marked tendency to vascular invasion involving the following sequential events: tumor cell contact with vascular BM, BM degradation possibly by 72 kDa gelatinases and endothelial disruption. This model could be a useful tool for understanding the mechanism of hematogenous metastasis of human gastric cancer.

Tumor-infiltrating lymphocytes (TIL) isolated from 11 gastric carcinoma were studied. TIL could grow for a long-term in medium containing recombinant interleukin-2(rIL-2). The mean expansion fold achieved in 6 long-term cultures of 11 specimens was 15.1 RIL-2 expanded gastric TIL exhibited significant cytotoxicity against K562, BGC823, MCF-7 and more effective antitumor cytotoxicity against fresh autologous tumor targets and human gastric cancer cell line. Peak cytotoxicity was shown in the third or fourth week after cultures. Cryopreservation of gastric TIL didn’t influence their expansion capacity and antitumor activity. Phenotypic analysis was demonstrated in this study. The results of present study indicate that TIL from human gastric carcinoma could be expanded and reach high levels of antitumor effector function in long-term cultures with rIL-2. Their function may be of clinical importance.

To explore the effect of intratumoral expressions of interleukin-12 (IL-12) and interleukin-18 (IL-18) on clinical features, angiogenesis and prognosis of gastric carcinoma.The expressions of IL-12 and IL-18 from 50 samples of gastric cancer tissue were analyzed by immunohistochemistry, and microvessel density (MVD) was determined with microscopic imaging analysis system.The positive expression rates of IL-12 and IL-18 were 44% (22/50) and 26% (13/50), respectively. IL-12 was significantly associated with pathologic differentiation, depth of invasion, lymph node metastasis, distant metastasis, and TNM stage, and IL-18 was closely related to distant metastasis. Intratumoral IL-12 and IL-18 expressions were not statistically related to MVD scoring. IL-12-positive patients survived significantly longer than those with IL-12-negative tumors, but there was no significant difference between IL-18-positive patients and IL-18-negative ones. The multivariate analysis with Cox proportional hazard model revealed IL-12, MVD and T stage were independent prognostic factors.The positive expressions of IL-12 and IL-18 can play an important role in progression and metastasis of gastric cancer, and IL-12 might be an independent factor of poor prognosis in gastric carcinoma.

75 Background: Gastric cancer (GAC) is the second most common cancer worldwide and the fifth leading cause of cancer-related death in Taiwan. To improve the survival of gastric cancer patients, biomarkers for early detection and effective anticancer therapy are required. Methods: Dysregulated proteins in GAC tumor cells were captured and identified by laser capture microdissection (LCM) and an isobaric tags for relative and absolute quantitation (iTRAQ)-labeling following basic-RP HPLC coupled to LTQ-orbitrap MS analysis. DEK, one of the most highly-expressed proteins in both intestinal and diffuse gastric cancers in iTRAQ analysis, was selected for further study. DEK proto-oncogene is originally identified as a fusion with the CAN nucleoporin protein in a subset of acute myeloid leukemia (AML) patients carrying the t(6;9) translocation, and DEK has been implicated in acute myeloid leukemia, melanoma, glioblastoma, hepatocellular carcinoma, and bladder cancer. The overexpression of DEK was confirmed using real-time quantitative-reverse transcription-polymerase chain reaction (Q-RT–PCR), Western blot, and immunohistochemical (IHC) analysis. Results: The data revealed that the expression of DEK mRNA was up-regulated in tumor tissues (compared with adjacent non-tumor tissues) of about 55.1% gastric patients. The expression of DEK protein was also up-regulated in tumor tissues identified by Western blot and IHC. To further understand the role of DEKgene in the human gastric cancer cells, the DEK knockdown stable clones were prepared in AZ521. The cells migration ability was reduced in DEK depletion stable clones, and the antitumor drug sensitivity and anoikis were increased in DEK depletion cells. Conclusions: These results indicate that DEK overexpression may contribute the poor prognosis of patients.

EBV infection has been causally associated with incidence of many carcinomas. EBV-associated gastric carcinoma (EBVaGC) has been classified as a unique gastric carcinoma subset, suggesting EBV infection is related to the development of gastric cancer. In this study, general trends of EBVaGC studies for last half-decades were reviewed in several perspectives of clinical significance, virological importance and etiological interests. Throughout this comprehensive reviewing, new study trends of EBV and EBVaGC for next half-decades were suggested.