miR-204抑制感染幽门螺杆菌的胃上皮细胞迁移和侵袭的机制研究

Mechanism of miR-204 inhibiting migration and invasion of gastric epithelial cells infected with Helicobacter pylori

目的探究miR-204抑制感染幽门螺杆菌(H.pylori)的胃上皮细胞迁移和侵袭的机制。方法将人胃上皮细胞GES1分为3组:对照组、H.pylori组和H.pylori+mimic组。H.pylori组和H.pylori+mimic组细胞按照100∶1的感染比例与H.pylori进行共培养。H.pylori+mimic组细胞在与H.pylori共培养之前转染miR-204 mimic以过表达miR-204。显微镜观察共培养情况。qPCR检测miR-204水平。细胞划痕实验和Transwell检测细胞迁移和侵袭能力。Western blotting检测EMT相关蛋白。结果感染H.pylori后,GES1细胞中的miR-204表达水平比正常GES1细胞显著降低(t=27.962,P=0.000)。H.pylori组的miR-204水平显著低于对照组(P<0.05),H.pylori+mimic组的miR-204水平显著高于对照组和H.pylori组(P<0.05)。H.pylori组的细胞迁移和侵袭能力显著高于对照组(P<0.05),H.pylori+mimic组的迁移和侵袭能力显著低于H.pylori组(P<0.05)。H.pylori组的E-cadherin蛋白显著高于对照组,而N-cadherin和Vimentin蛋白显著低于对照组(P<0.05)。H.pylori+mimic组的E-cadherin显著低于H.pylori组而N-cadherin和Vimentin显著高于H.pylori组(P<0.05)。结论将胃上皮细胞与H.pylori共培养会使细胞中miR-204的水平降低,且过表达miR-204会通过调节上皮-间充质转化抑制由H.pylori引起的胃上皮细胞迁移和侵袭。

Objective To investigate the mechanism of miR-204 inhibiting the migration and invasion of gastric epithelial cells infected with H.pylori.Methods Human gastric epithelial cells GES1 were divided into three groups:control group,H.pylori group,and H.pylori+mimic group.The cells of H.pylori group and H.pylori+mimic group were cocultured with H.pylori at the ratio of 100∶1.H.pylori+mimic group cells were transfected with miR-204 mimic before co-culture with H.pylori to overexpress miR-204.Co-culture was observed under a microscope.qPCR was used to detect miR-204 levels.Cell scratch test and Transwell was applied to detect cell migration and invasion.Western blotting was used to detect EMT-related proteins.Results After infection with H.pylori,the level of miR-204 in GES1 cells was lower than that in normal GES1 cells(t=27.962,P=0.000).The level of miR-204 in the H.pylori group was significantly lower than that in the control group(P<0.05).The levels of miR-204 in H.pylori+mimic group were significantly higher than those in the control group and H.pylori group(P<0.05).The cell migration and invasion ability of the H.pylori group was significantly higher than those of the control group(P<0.05).The migration and invasion ability of the H.pylori+mimic group was significantly lower than those of the H.pylori group(P<0.05).The E-cadherin protein in the H.pylori group was significantly higher than that in the control group,while the N-cadherin and Vimentin proteins were significantly lower than those in the control group(P<0.05).E-cadherin in the H.pylori+mimic group was significantly lower than that in the H.pylori group,while N-cadherin and Vimentin were significantly higher than those in the H.pylori group(P<0.05).Conclusion Co-culture of gastric epithelial cells with H.pylori will reduce the levels of miR-204 in the cells,and overexpression of miR-204 will inhibit the migration and invasion of gastric epithelial cells caused by H.pylori by regulating epithlial mesenchymal transition.