Tip30基因过表达对人胃癌细胞生长的影响及分子机制

The effect and molecular mechanism of Tip30 gene overexpression on cell growth in human gastric cancer cells

目的探讨Tip30基因过表达对人胃癌AGS细胞增殖、凋亡的影响及相关分子机制。方法人胃癌AGS细胞中Tip30基因过表达采用腺病毒转染法,使用实时定量PCR和Western blotting验证Tip30过表达的效果,分别采用MTT法、划痕试验检测细胞增殖、迁移情况,流式细胞术分析细胞凋亡,Western blotting分析p53、Bax、Bcl-2蛋白表达。结果过表达组AGS细胞Tip30基因m RNA和蛋白表达水平均显著高于对照组(P<0.05)。与对照组比较,Tip30过表达后,细胞增殖、体外迁移能力均显著减弱(P<0.05);Tip30过表达后AGS细胞凋亡率相比对照组显著增大,Tip30过表达促进AGS细胞凋亡与上调p53、Bax蛋白表达,下调Bcl-2蛋白表达水平相关。结论 Tip30基因过表达可抑制人胃癌AGS细胞增殖活性、体外迁移能力,且促进胃癌细胞凋亡。

Objective To investigate the effect and underlying molecular mechanism of Tip30 gene overexpression on cell proliferation and apoptosis in human gastric cancer AGS cells. Methods Adenovirus transfection was used to overexpress the Tip30 gene in human gastric cancer AGS cells. Tip30 overexpression was verified by real-time PCR and western blot. Cell proliferation was analyzed with the MTT method, a wound scratch assay was used to monitor cell migration activity, and the cell apoptosis rate was detected with flow cytometry. The expression level of p53, Bax and Bcl-2 was detected with western blot. Results The m RNA and protein levels of Tip30 in human gastric cancer AGS cells were significantly higher in the overexpression group compared with the control group（P0.05）. The proliferation and migratory ability of AGS cells in vitro were decreased when Tip30 was overexpressed（P0.05）. The cell apoptosis rate was significantly increased in Tip30 overexpressing cells, and a corresponding up-regulation of p53 and Bax and down-regulation of Bcl-2 was also observed. Conclusion Overexpression of Tip30 in human gastric cancer AGS cells inhibits both cell proliferation and migration in vitro, and enhances the AGS cell apoptosis.