雷公藤红素通过PI3K/AKT/mTOR信号通路对胃癌MFC细胞增殖和凋亡的影响

Effects of tripterine on the proliferation and apoptosis of gastric cancer MFC cells by regulating PI3K/AKT/mTOR signaling pathway

目的研究雷公藤红素对胃癌MFC细胞内PI3K/AKT/mTOR信号通路的影响及其对胃癌MFC细胞增殖的抑制和促凋亡作用机制。方法分别设置对照组和雷公藤红素低、中、高剂量(5、10、20 mmol/L)组,采用MTT法检测雷公藤红素对胃癌MFC细胞增殖的影响;采用流式细胞技术检测雷公藤红素对胃癌MFC细胞周期的影响及MFC细胞凋亡的情况;Western blotting检测雷公藤红素对胃癌MFC细胞内凋亡相关蛋白Bax、Bcl-2和PI3K、AKT和mTOR蛋白表达的影响;实时定量PCR检测雷公藤红素对胃癌MFC细胞内PI3K、AKT和mTOR mRNA表达的影响。结果与对照组比较,雷公藤红素能够呈剂量相关性地显著抑制胃癌MFC细胞的增殖(P<0.05);与对照组比较,雷公藤红素能够呈剂量相关性地显著引起MFC细胞G2/M期阻滞(P<0.05);与对照组比较,雷公藤红素能够呈剂量相关性地显著促进MFC细胞的凋亡(P<0.05);与对照组比较,雷公藤红素能够呈剂量相关性地显著抑制MFC细胞内抗凋亡蛋白Bcl-2蛋白和PI3K、AKT、mTOR蛋白的表达(P<0.05),显著促进MFC细胞内促凋亡蛋白Bax的表达(P<0.05);与对照组比较,雷公藤红素能够呈剂量相关性地显著抑制MFC细胞内PI3K、AKT和mTOR mRNA的表达(P<0.05)。结论雷公藤红素能够抑制胃癌MFC细胞的增殖并促进胃癌MFC细胞凋亡,其作用机制可能是通过抑制胃癌MFC细胞内PI3K/AKT/mTOR信号通路中PI3K、AKT、mTOR等蛋白的表达而抑制胃癌MFC细胞的增殖,进而促进胃癌MFC细胞内促凋亡蛋白Bax的表达,同时抑制抗凋亡蛋白Bcl-2的表达,从而促进胃癌MFC细胞凋亡。

Objective To study the effect of tripterine on PI3K/AKT/mTOR signaling pathway in gastric cancer MFC cells and its mechanism of inhibiting proliferation and promoting apoptosis of gastric cancer MFC cells.Methods The control group,low dose medium dose,and high dose(5,10,20 mmol/L)groups were used to test the proliferation of gastric cancer MFC cells.MTT assay was used to detect the effects of tripterine on the proliferation of gastric cancer MFC cells.The effects of tripterine on the cell cycle of gastric cancer MFC cells and the apoptosis of MFC cells were detected by flow cytometry.The effects of tripterine on apoptosis-related proteins Bax,Bcl-2,and the expression of PI3K,AKT and mTOR proteins in gastric cancer MFC cells were studied by Western blotting.The effects of tripterine on the expression levels of PI3K,AKT and mTOR mRNA in gastric cancer MFC cells were detected by real-time quantitative PCR.Results Compared with the control group,tripterine could significantly inhibit the proliferation of gastric cancer MFC cells in a dose-dependent manner(P<0.05).Compared with the control group,tripterine could significantly induce G2/M phase arrest of MFC cells in a dose-dependent manner(P<0.05).Compared with the control group,tripterine could significantly promote the apoptosis of MFC cells in a dose-dependent manner(P<0.05).Compared with the control group,tripterine could significantly inhibit the expression of anti-apoptotic protein Bcl-2,and PI3K,AKT,and mTOR proteins in MFC cells in a dose-dependent manner(P<0.05),but promote the expression of pro-apoptotic protein Bax in MFC cells in a dose-dependent manner(P<0.05).Compared with the control group,tripterine could significantly inhibit the transcription of PI3K,AKT and mTOR mRNA in MFC cells in a dose-dependent manner(P<0.05).Conclusion Tripterine can inhibit the proliferation of gastric cancer MFC cells and promote the apoptosis of gastric cancer MFC cells.The mechanism of action may be to inhibit the proliferation of gastric cancer MFC cells by inhibiting the expression of PI3K,AKT,mTOR and other proteins in PI3K/AKT/mTOR signaling pathway in gastric cancer MFC cells,and then promote the expression of apoptosis-promoting protein Bax in gastric cancer MFC cells.At the same time,it can inhibit the expression of anti-apoptotic protein Bcl-2 and promote the apoptosis of gastric cancer MFC cells.