Effect of Helicobacter pylori on apoptosis and apoptosis related genes in gastric cancer cells.

Y Yang, C S Deng, J Z Peng, B C-Y Wong, S K Lam, H H-X Xia
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Abstract

Background/aims: Helicobacter pylori induces the apoptosis of gastric epithelial cells in vivo and in vitro. However, the molecular mechanism has not been clarified. The aim of this study was to investigate the effect of H pylori on the apoptosis of gastric epithelial cells and the expression of apoptosis related genes in vitro.

Methods: Human gastric adenocarcinoma SGC-7901 cells were co-cultured with a cytotoxic H pylori strain, NCTC 11637, at various densities ranging from 3.2 x 10(4) to 1.0 x 10(8) colony forming units (CFU)/ml for 48 hours. Apoptosis in gastric cells was determined by transmission electron microscopy, Hoechst 33258 fluorochrome staining, and flow cytometry. The expression of apoptosis related proteins, Bcl-2, Bax, and c-Myc, was measured by an immunohistochemical method, and c-Myc mRNA expression was determined by the reverse transcription-polymerase chain reaction.

Results: Helicobacter pylori induces morphological changes typical of apoptosis. Both fluorochrome staining and flow cytometry showed that the apoptotic index began to increase when H pylori were at a density of > 1.6 x 10(4) CFU/ml, and in a density dependent manner (p < 0.01; one way ANOVA). The expression of the Bax and c-Myc proteins and of c-Myc mRNA was increased, whereas Bcl-2 expression was decreased after co-culture for 48 hours.

Conclusions: Helicobacter pylori induced apoptosis in gastric epithelial cells is mediated by altered expression of the products of the Bcl-2, Bax, and c-Myc genes.

幽门螺杆菌对胃癌细胞凋亡和凋亡相关基因的影响
背景/目的:幽门螺杆菌可诱导体内和体外胃上皮细胞凋亡。然而,其分子机制尚未明确。方法:将人胃腺癌 SGC-7901 细胞与细胞毒性幽门螺杆菌菌株 NCTC 11637 以 3.2 x 10(4) 到 1.0 x 10(8) 菌落总数形成单位 (CFU)/ml 的不同密度共培养 48 小时。通过透射电子显微镜、Hoechst 33258 荧光染色和流式细胞术检测胃细胞的凋亡情况。免疫组化法测定了凋亡相关蛋白 Bcl-2、Bax 和 c-Myc 的表达,反转录聚合酶链反应测定了 c-Myc mRNA 的表达:结果:幽门螺杆菌可诱导典型的细胞凋亡形态学变化。荧光染色法和流式细胞术均显示,当幽门螺杆菌的密度大于 1.6 x 10(4) CFU/ml 时,细胞凋亡指数开始增加,且呈密度依赖性(p < 0.01;单向方差分析)。共培养 48 小时后,Bax 和 c-Myc 蛋白以及 c-Myc mRNA 的表达增加,而 Bcl-2 的表达减少:结论:幽门螺杆菌诱导的胃上皮细胞凋亡是由 Bcl-2、Bax 和 c-Myc 基因产物表达的改变介导的。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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