胃癌平诱导人胃癌细胞株SGC-7901凋亡的实验研究

目的:探讨胃癌平对诱导人胃癌细胞株SGC-7901凋亡的影响。方法:采用体外培养胃癌细胞株SGC-7901,MTT法检测胃癌细胞增殖抑制率、观察光镜下形态变化及电镜下胃癌细胞株SGC-7901凋亡情况。结果:与对照组相比胃癌平方各浓度组、DDP组以及中西药结合组24h肿瘤细胞增殖抑制率均有统计学的意义(P<0.01)。光镜下观察胃癌平方组细胞失去正常生长形态,长梭形细胞变圆,胞浆浓缩,核浓缩。电镜下可见凋亡小体。结论:胃癌平方具有抑制人胃癌细胞株SGC-7901的增殖、诱导凋亡,可能是该药抗肿瘤机制之一。

消癌平片在控制胃癌患者化疗期间急性不良反应中的作用

目的探讨消癌平片在控制胃癌患者化疗期间急性不良反应的临床效果。方法将我院自2011年12月至2015年12月间收治的胃癌根治患者106例作为研究对象,随机的分为研究组和对照组各53例,两组患者均采用常规化疗,研究组在此基础上加服消癌平片,观察两组患者化疗期间急性不良反应发生率和患者生存治疗。结果研究组患者恶心呕吐、脱发、肝功能异常、白细胞降低、感染性发热等急性不良反应发生率显著低于对照组,术后1个月、3个月、6个月KPS评分均显著高于同期对照组(P<0.05),组间比较具有统计学意义。结论行胃癌根治术患者化疗期间辅助使用消癌平片能够减少急性不良反应发生率,促进患者康复,提高患者生存质量,效果显著,可在临床范围内推广使用。

Relationship between LYVE-1, VEGFR-3 and CD44 gene expressions and lymphatic metastasis in gastric cancer

AIM: To investigate the expression levels of lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1), vascular endothelial growth factor receptor-3 (VEGFR-3) and CD44 genes and the relationship between their lev- els and clinicopathological parameters in gastric cancer.METHODS: Tissue samples were obtained from 33 patients (8 females) with gastric cancer. mRNA levels of LYVE-1, VEGFR-3 and CD44 in normal and tumor tissues were quantitatively measured using real time polymerase chain reaction. The results were correlated with lymph node metastasis, histological type and differentiation of the tumor, T-stage, and presence of vascular, perineural and lymphatic invasions. The distribution of molecules in the tissue was evaluated using immunohistochemistry. RESULTS: LYVE-1, CD44 and VEGFR-3 gene expression levels were significantly higher in gastric cancer than in normal tissue. While there was no correlation between gene expressions and clinicopathologic fea- tures such as histologic type, differentiation and stage, gene expression levels were found to be increased in conjunction with positive lymph node/total lymph node ratio and the presence of perineural invasion. A significant correlation was also found between LYVE-1 and CD44 over-expressions and perineural invasion and lymph node positivity in gastric cancers. When the dis- tribution of LYVE-1 antibody-stained lymphatic vessels in tissue was evaluated, lymphatic vessels were located intra-tumorally in 13% and peri-tumorally in 27% of the patients. Moreover, lymph node metastases were also positive in all patients with LYVE-1-staining. CONCLUSION: LYVE-1, VEGFR-3 and CD44 all play an important role in lymphangiogenesis, invasion and metastasis. LYVE-1 is a perfectly reliable lymphatic vessel marker and useful for immunohistochemistry.

The relationship between level of plasma D-dimer and survival time in metastatic gastric cancer patients

Objective:The principal purpose of this study was to determine the relationship between level of plasma D-dimer and survival time in metastatic gastric cancer patients.Methods:We retrospectively collected the data of plasma D-dimer in metastatic gastric cancer patients admitted in our Department (Department of Oncology,The Affiliated Changzheng Hospital,The Second Military Medical University,Shanghai,China) from October 2006 to October 2008 and analyzed the relationship between level of plasma D-dimer and survival time along with other clinicopathologic parameters.Results:A total of 82 patients were studied in our research,52 were males and 30 females,and the mean age was 57 years.The 48 cases had a normal plasma D-dimer level (<300μg/L) and 34 had a high plasma D-dimer level (≥300μg/L).In the normal and high plasma D-dimer level groups,the mean survival times were 10.9 (95% CI:9.8-12.2) months and 6.8(95% CI:4.4-7.6) months respectively,and the difference was statistically significant.Conclusion:Metastatic gastric cancer patients with high plasma D-dimer level had significantly shorter survival time than those with normal plasma D-dimer level.Level of plasma D-dimer can be referred as a potential predictor in metastatic gastric cancer patients.

FAT10 level in human gastric cancer and its relation with mutant p53 level,lymph node metastasis and TNM staging

AIM:To investigate the role of FAT10 and mutant p53 in the pathogenesis,severity and prognosis of gastric cancer.METHODS:FAT10,mutant p53 mRNA and protein levels were measured by reverse transcription(RT)-PCR and immunohistochemistry in gastric cancer tissue(n = 62),tumoradjacent tissue(n = 62) and normal gastric tissue(n = 62).Relation of FAT10 and mutant p53 expression with clinicopathological features and clinical outcomes of gastric cancer patients were analyzed.RESULTS:The FAT10,mutant p53 mRNA and protein levels were signif icantly higher in gastric cancer than in its adjacent and normal tissue.The FAT10 and mutant p53 levels in gastric cancer tissue were significantly correlated with lymph node metastasis and tumor,nodes,metastasis(TNM) staging.Moreover,the high FAT10 level was associated with the overall survival rate of patients.Multivariate Cox-proportional hazards model analysis showed that mRNA and protein levels of FAT10 and mutant p53,lymph node metastasis,distant metastasis and TNM stage were the independent prognostic factors for gastric cancer.CONCLUSION:FAT10 may be involved in gastric carcinogenesis,and is a potential marker for the prognosis of gastric cancer patients.FAT10 and mutant p53 may play a common role in the carcinogenesis of gastric cancer.

Clinical significance of human kallikrein 12 gene expression in gastric cancer

AIM:To investigate whether the expression of kallikrein 12(KLK12) is related to the development of gastric cancer(GC) and to determine the role of KLK12 in gastric cancer cells growth,invasion and migration.METHODS:Between September 2007 and March 2008,133 patients with histologically confirmed GC were recruited for the study.Expression of KLK12 was detected in samples from GC patients by quantitative real-time reverse transcription polymerase chain reaction and immunohistochemistry.The relationship between KLK12 protein expression and clinicopathological features of GC was analyzed.The difference in 5-year survival rates between the high KLK12 protein expression group and the low KLK12 expression group was compared.Additionally,the expression of KLK12 was examined in various human GC cell lines,including MKN-28,SGC-7901 and MKN-45.Small interfering RNA(siRNA) was used to inhibit KLK12 expression in MKN-45 cells.Cell clones stably transfected with KLK12 siRNA were tested for KLK12 expression by quantitative real-time reverse transcription-polymerase chain reaction and Western blotting.Furthermore,a series of functional assays were performed in this study to assess the biological features of transfected cells.Cell proliferation was assessed using the methylthiazolyltetrazoliumassay.Finally,cell migration and invasion were assessed using transwell chamber assays.RESULTS:Of the 133 GC patients included in the study,126(94.7%) showed a higher expression level of KLK12 mRNA when compared to noncancerous tissue specimens.Expression of KLK12 mRNA was significantly higher in GC tissues than in normal tissue(P < 0.001).KLK12 protein expression was detected in 96 of 133(72.2%) GC samples with moderate or strong staining primarily in the cytoplasm.In contrast,negative immunostaining for KLK12 protein was observed in the corresponding normal gastric mucosal tissue.Overexpression of KLK12 protein was significantly associated with lymph node metastasis(P = 0.001),histological type(P < 0.001) and tumor-node-metastasis stage(P = 0.005),while no significant correlation was observed between expression of KLK12 protein and sex,age,depth of invasion,tumor size or lymphatic invasion.Furthermore,patients with high KLK12 expression had a significantly poorer 5-year survival rate than those with low KLK12 expression(P = 0.002).Expression of KLK12 mRNA was significantly higher in MKN-45 GC cells compared to normal mucosal cells or two other GC cell lines(P < 0.01).Expression of KLK12 in MKN-45 cells was downregulated after transfection with siRNA.Knockdown of KLK12 markedly decreased the proliferation of MKN-45 cells when compared with parent or mock-transfected cells(P = 0.001),especially from the 3rd to the 5th day of the assay.In migration assays,fewer KLK12 siRNA cells migrated through the chambers(22.00 ± 1.81) when compared to the parent(46.47 ± 2.42) or mock-transfected cells(45.40 ± 1.99);these differences were statistically significant(P < 0.001).However,in the invasion assay,the number of KLK12 siRNA cells that invaded the chambers was 18.40 ± 1.12,closely similar to both the parent(18.67 ± 0.98) and mock-transfected cells(18.53 ± 0.92).There was no significantly difference between the three groups in the invasion assay(P = 0.054).CONCLUSION:The KLK12 gene is markedly overexpressed in GC tissue,and its expression status may be a powerful prognostic indicator for patients with GC.KLK12 might serve as a novel diagnosis and prognosis biomarker in GC.

Effects of vagus nerve preservation and vagotomy on peptide YY and body weight after subtotal gastrectomy

AIM:To investigate the relationship between the function of vagus nerve and peptide YY 3-36 and ghrelin levels after subtotal gastrectomy.METHODS:We enrolled a total of 16 patients who underwent subtotal gastrectomy due to gastric cancer.All surgeries were performed by a single skilled surgeon.We measured peptide YY 3-36,ghrelin,leptin,insulin,growth hormone levels,and body weight immediately before and one month after surgery.RESULTS:Vagus nerve preservation group showed less body weight loss and less increase of peptide YY 3-36 compared with vagotomy group(-5.56 ± 2.24 kg vs-7.85 ± 1.57 kg,P = 0.037 and 0.06 ± 0.08 ng/mL vs 0.19 ± 0.12 ng/mL,P = 0.021,respectively).Moreover,patients with body weight loss of less than 10% exhibited reduced elevation of peptide YY 3-36 level,typically less than 20% [6(66.7%) vs 0(0.0%),P = 0.011,odd ratio = 3.333,95% confidence interval(1.293,8.591)].CONCLUSION:Vagus nerve preservation contributes to the maintenance of body weight after gastrectomy,and this phenomenon may be related to the suppressed activity of peptide YY 3-36.

Decreased blood riboflavin levels are correlated with defective expression of RFT2 gene in gastric cancer

AIM:To investigate the relationship between blood riboflavin levels and riboflavin transporter 2(RFT2) gene expression in gastric carcinoma(GC) development.METHODS:High-performance liquid chromatography was used to detect blood riboflavin levels in patients with GC.Real-time fluorogenic quantitative polymerase chain reaction and immunohistochemistry were used to analyze the expression of RFT2 mRNA and protein in samples from 60 GC patients consisting of both tumor and normal tissue.RESULTS:A significant decrease in the RFT2 mRNA levels was detected in GC samples compared with those in the normal mucous membrane(0.398 ± 0.149 vs 1.479 ± 0.587;P = 0.040).Tumors exhibited low RFT2 protein expression(75%,16.7%,8.3% and 0% for no RFT2 staining,weak staining,medium staining and strong staining,respectively),which was significantly lower than that in the normal mucous membrane(10%,16.7%,26.7% and 46.7% for no RFT2 staining,weak staining,medium staining and strong staining,respectively;P < 0.05).Tumors with low RFT2 expression were significantly associated with tumor stage and histological grade.Moreover,a significantly decrease in Uyghur patients was observed compared with Han patients.However,other parameters-gender,tumor location and lymph node metastasis-showed no significant relationship with RFT2 expression.Blood riboflavin levels were reverse correlated with development of GC(1.2000 ± 0.97 569 ng/mL in high tumor stage patients vs 2.5980 ± 1.31 129 ng/mL in low tumor stage patients;P < 0.05).A positive correlation of plasma riboflavin levels with defective expression of RFT2 protein was found in GC patients(2 = 2.619;P = 0.019).CONCLUSION:Defective expression of RFT2 is associated with the development of GC and this may represent a mechanism underlying the decreased plasma riboflavin levels in GC.

Gastric choriocarcinoma admixed with an α-fetoprotein-producing adenocarcinoma and separated adenocarcinoma

We report a case of gastric choriocarcinoma admixed with an α-fetoprotein (AFP)-producing adeno-carcinoma. A 70-year-old man was hospitalized for gastric cancer that was detected during screening by esophagogastroduodenoscopy (EGD). Initial laboratory data showed the increased serum level of AFP and EGD revealed a 5-cm ulcerofungating mass in the greater curvature of the gastric antrum. The patient underwent radical subtotal gastrectomywith D2 lymph node dissection and Billroth gastrojejunostomy. Histopathological evaluation confirmed double primary gastric cancer: gastric choriocarcinoma admixed with an AFP-producing adenocarcinoma and separated adenocarcinoma. At 2 wk postoperatively, his human chorionic gonadotropin and AFP levels had reduced and six cycles of adjuvant chemotherapy were initiated. No recurrence or distant metastasis was observed at 4 years postoperatively.

Overexpression of the M2 isoform of pyruvate kinase is an adverse prognostic factor for signet ring cell gastric cancer

AIM:To investigate M2 isoform of pyruvate kinase(PKM2) expression in gastric cancers and evaluate its potential as a prognostic biomarker and an anticancer target.METHODS:All tissue samples were derived from gastric cancer patients underwent curative gastrectomy as a primary treatment.Clinical and pathological information were obtained from the medical records.Gene expression microarray data from 60 cancer and 19 noncancer gastric tissues were analyzed to evaluate the expression level of PKM2 mRNA.Tissue microarrays were constructed from 368 gastric cancer patients.Immunohistochemistry was used to measure PKM2 expression and PKM2 positivity of cancer was determined by proportion of PKM2-positive tumor cells and staining intensity.Association between PKM2 expression and the clinicopathological factors was evaluated and the correlation between PKM2 and cancer prognosis was evaluated.RESULTS:PKM2 mRNA levels were increased more than 2-fold in primary gastric cancers compared to adjacent normal tissues from the same patients(log transformed expression level:7.6 ± 0.65 vs 6.3 ± 0.51,P < 0.001).Moreover,differentiated type cancers had significantly higher PKM2 mRNA compared to undifferentiated type cancers(log transformed expression level:7.8 ± 0.70 vs 6.7 ± 0.71,P < 0.001).PKM2 protein was mainly localized in the cytoplasm of primary cancer cells and detected in 144 of 368(39.1%) human gastric cancer cases.PKM2 expression was not related with stage(P = 0.811),but strongly correlated with gastric cancer differentiation(P < 0.001).Differentiated type cancers expressed more PKM2 protein than did the undifferentiated ones.Well differentiated adenocarcinoma showed 63.6% PKM2-positive cells;in contrast,signet-ring cell cancers showed only 17.7% PKM2-positive cells.Importantly,PKM2 expression was correlated with shorter overall survival(P < 0.05) independent of stage only in signet-ring cell cancers.CONCLUSION:PKM2 expression might be an adverse prognostic factor for signet-ring cell carcinomas.Its function and potential as a prognostic marker should be further verified in gastric cancer.

CXCR4/SDF-1 axis is involved in lymph node metastasis of gastric carcinoma

AIM:To investigate the role of CXC chemokine receptor-4 (CXCR4) and stromal cell-derived factor-1 (SDF-1) in lymph node metastasis of gastric carcinoma.METHODS:In 40 cases of gastric cancer,expression of CXCR4 mRNA in cancer and normal mucous membrane and SDF-1 mRNA in lymph nodes around the stomach was detected using quantitative polymerase chain reaction (PCR) (TaqMan) and immunohistochemistric assay.SGC-7901 and MGC80-3 cancer cells were used to investigate the effect of SDF-1 on cell proliferation and migration.RESULTS:Quantitative reverse transcription PCR and immunohistochemistry revealed that the expression level of CXCR4 in gastric cancer was significantly higher than that in normal mucous membrane (1.6244 ± 1.3801 vs 1.0715 ± 0.5243,P < 0.05).The expression level of CXCR4 mRNA in gastric cancer with lymph node metastasis was also significantly higher than that without lymph node metastasis (0.823 ± 0.551 vs 0.392 ± 0.338,P < 0.05).CXCR4 expression was significantly related to poorly differentiated,high tumor stage and lymph node metastasis.Significant differences in the expression level of SDF-1 mRNA were found between lymph nodes in metastatic gastric cancer and normal nodes (0.5432 ± 0.4907 vs 0.2640 ± 0.2601,P < 0.05).The positive expression of SDF-1 mRNA in lymph nodes of metastatic gastric cancer was consistent with the positive expression of CXCR4 mRNA in gastric cancer (r=0.776,P < 0.01).Additionally,human gastric cancer cell lines expressed CXCR4 and showed vigorous proliferation and migratory responses to SDF-1.AMD3100 (a specific CXCR4 antagonist) was also found to effectively reduce the migration of gastric cancer cells.CONCLUSION:The CXCR4/SDF-1 axis is involved in the lymph node metastasis of gastric cancer.CXCR4 is considered as a potential therapeutic target in the treatment of gastric cancer.

Effect of low-dose X-ray radiation on adaptive response in gastric cancer cell

Objective： We aimed to study the effect and mechanism of low-dose radiation （LDR） on adaptive response of gastric cancer cell. Methods： SGC7901 cells were cultured in vitro, and divided into 4 groups： control group （DO group）, low-dose radiation group （D1 group, 75 mGy）, high-dose radiation group （D2 group, 2 Gy）, low-dose plus high-dose radiation group （D1 ＋ D2 group, 75 mGy ＋ 2 Gy, the interval of low and high-close radiation being 8 h）. Cell inhibition rate was detected by cytometry and CCK8 method; the proportion of cell cycle at different times after irradiation was determined by using a flow cytometry. The ATM mRNA levels were detected by using quantitative real-time reverse transcription polymerase chain reaction （RT-PCR）. Results： There was no significant different between groups DO and D1, groups D2 and D1 ＋ D2 cell inhibition rate （P 〉 0.05）. There was a significant increase G2/M arrest in groups D2 and D1 ＋ D2 than groups DO and D1 after 6 h of radiation and did not recover at 48 h （P 〈 0.05）. The ATM mRNA expression of group D2 and D1 ＋ D2 increased highly than that of group DO and D1 （P 〈 0.05）. However, differences between group D2 and D1 ＋ D2, group DO and D1 were not statistical significant （P 〉 0.05）. Conclusion： LDR cannot induce adaptive response in SGC7901 cells in vitro, which may be associated the regulation of cell cycle, and its ATM mRNA expression cannot be affected by 75 mGy X-ray radiation.