幽门螺杆菌诱导胃癌细胞上皮-间充质转化的研究

Helicobacter pylori induces the epithelial-mesenchymal transition of gastric cancer cells

目的探讨幽门螺杆菌对胃癌细胞上皮-间充质转化(EMT)的诱导作用。方法设置空白对照组、实验组和阳性对照组,分别以幽门螺杆菌SS1菌株和转化生长因子-β(TGF-β)刺激胃黏膜上皮GES-1细胞、胃癌SGC-7901细胞。采用Transwell小室检测细胞纵向迁移能力;Western blot法测定EMT相关蛋白的表达变化规律;荧光共聚焦显微镜观察β-catenin的分布变化。结果 Transwell实验结果显示经幽门螺杆菌SS1菌株刺激后,GES-1细胞和SGC-7901细胞的迁移数目明显多于空白对照组,少于阳性对照组。Western blot显示幽门螺杆菌SS1菌株能抑制GES-1细胞和SGC-7901细胞E-钙黏蛋白(E-cadherin)的表达,上调SGC-7901细胞N-cadherin、Twist和基质金属蛋白酶-9(MMP-9)的表达,但对GES-1细胞N-cadherin和Vimentin的表达水平无明显影响。免疫荧光结果表明未接受刺激的两种细胞的β-catenin分布于胞质,而接受幽门螺杆菌SS1菌株刺激后β-catenin向细胞核移位。结论幽门螺杆菌能诱导胃癌SGC-7901细胞发生EMT,促进胃癌细胞迁移。

Objective To investigate the effect of Helicobacter pylori(H.pylori)on epithelial-mesenchymal transition(EMT)in gastric cancer cells.Methods Three groups were set,including the blank control group,experimental group and positive control group.The GES-1 cells and gastric cancer SGC-7901 cells were stimulated with H.pylori SS1 strain and TGF-β,respectively.The ability of cellular longitudinal migration was detected by Transwell assays.Western blot was used to examine the change rule of EMT-associated protein expression.Moreover,the distributions ofβ-catenin was observed by fluorescence confocal microscope.Results The Transwell assays results indicated that the migration number of GES-1 cells and SGC-7901 cells after SSI strain stimulation were significantly more than that of the blank control group,and less than that of the TGF-βpositive control group.Western blot showed that H.pylori SSI strain could inhibit the expression of E-cadherin in GES-1 cells and SGC-7901 cells,up-regulated the expression of N-cadherin,Twist and MMP-9 in SGC-7901 cells,while had no obvious effect on the expression of N-cadherin and Vimentin in GES-1 cells.The immunofluorescence assays results demonstrated thatβ-catenin in the two kinds of cells without receiving the stimulation were distributed in cytoplasm,whileβ-catenin had the translocation to the nucleus after receiving SSI strain stimulation.Conclusion H.pylori can induce the EMT occurrence in gastric cancer SGC-7901 cells,and enhances the migration of gastric cancer cells.