Effect of Helicobacter pylori on expression Levels of C-Met gene in HepG2 cells
陈光佩 1黎伟康 2戴佩玲 2吴绍峰 2杨巧红 3许晓伍 1刘坤东2
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作者信息
1. 广州中医药大学基础医学院人体解剖学系,广东 广州 510006
2. 广州中医药大学基础医学院实验教学中心,广东 广州 510006
3. 广州中医药大学基础医学院病理学与病理生理学系,广东 广州 510006
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摘要
目的 研究幽门螺杆菌(Hp)对肝癌HepG2细胞原癌基因C-Met表达的影响.方法 将Hp与HepG2细胞共同培养,设置空白对照组、Hp组、C-Met抑制剂空白组和C-Met抑制剂Hp组,每组设副孔5个,采用CCK8试剂盒检测HepG2细胞增殖活性,采用Western blot和qPCR方法检测C-Met原癌基因蛋白和mRNA表达水平.结果 相较于空白对照组,Hp处理组HepG2细胞增殖活性升高且与时间呈正相关,而C-Met抑制剂可以抑制HepG2细胞的增殖活性.同时,Hp组HepG2细胞C-Met基因蛋白和mRNA表达水平均上调,C-Met抑制剂组则显著下调.相较于Hp共培养12 h,共培养24 h C-Met蛋白和mRNA表达水平均上调.结论 Hp促进HepG2细胞的增殖可能与其提高原癌基因C-Met的蛋白及其mRNA表达水平有关,为后续更深入研究肝癌发生发展的复杂机制提供了理论依据.
Abstract
Objective To investigate the effects of Helicobacter pylori(Hp)on proto-oncogene C-Met in HepG2 cells.Methods Hp and HepG2 cells were co-cultured and the control group,HP group,c-Met inhibitor treatment group and c-Met inhibitor treatment Hp group were set up.Each group had 5 side holes.The proliferation activity of HepG2 cells was detected by CCK8 kit,and the expression level of C-Met protein and mRNA was detect-ed by Western blot and qPCR.Results Compared with the control group,the proliferation activity of HepG2 cells in Hp treatment group was increased and positive correlation with time,while the c-met inhibitor treatment group was down-regulated.Meanwhile,the expression levels of c-met protein and mRNA in Hp treatment group was ascended,while the c-met inhibitor treatment group decreased.Compared with 12 h,the expression levels of c-met protein and mRNA in Hp treatment for 24 h was up-regulated.Conclusion Hp can promote the proliferation of HepG2 cells,which may be related to the increase of the expression levels of proto-oncogene C-Met proteins and its mRNA in HepG2 cells,which provides a theoretical basis for further research on the complex mechanism of hepatocarcinogene-sis and development.
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