Akt抑制剂perifosine抑制胃癌细胞增殖并诱导凋亡的实验研究

Perifosine inhibits cell proliferation and triggers apoptosis in human gastric cancer cell line SGC-7901 cells

目的:探讨新的Akt抑制剂perifosine对胃癌细胞增殖与凋亡的影响。方法:采用MTT法检测perifosine对胃癌细胞SGC-7901细胞增殖的影响;流式细胞术分析细胞周期变化;Annexin Ⅴ-FITC试剂盒检测细胞凋亡;免疫印迹法检测蛋白表达水平。结果:Perifosine能够抑制SGC-7901细胞的增殖,且抑制作用呈时间和剂量依赖性。胃癌细胞经perifosine处理后,细胞阻滞于G2期,p21表达水平增加,而cyclin B1的表达受到抑制;凋亡诱导现象随着perifosine剂量的增加而增强。免疫印迹结果显示perifosine活化SGC-7901细胞内caspase-3﹑caspase-9及其底物多聚(ADP-核糖)聚合酶(PARP),促进凋亡诱导蛋白Bax的表达,并抑制Bcl-2的表达。结论:Perifosine对人胃癌细胞SGC-7901的生长具有显著的抑制作用,其凋亡诱导活性与调节caspase家族和Bcl-2家族蛋白的表达密切相关。

AIM：To investigate the effect of perifosine,a novel inhibitor of Akt,on the cell proliferation and apoptosis in human gastric cancer cell line SGC-7901.METHODS： Cell growth inhibition was detected by MTT assay.Cell cycle was analyzed by flow cytometry.Annexin V-FITC apoptosis detection kit was used to determine apoptosis in the cells.Protein expression was examined by Western blotting.RESULTS： Akt phosphorylation was dose-dependently inhibited by perifosine in SGC-7901 cells.The results of MTT and cell cycle analysis indicated that perifosine inhibited the growth of human gastric cancer cells in a dose-dependent manner.Perifosine arrested the cell cycle progression at G2 phase.Apoptosis induction became more effective with increasing the concentration of perifosine.The caspase cascade and its downstream effector poly（ADP-ribose） polymerase（PARP） were also activated upon perifosine treatment,and the level of Bcl-2 was down-regulated,whereas the protein level of Bax was up-regulated.CONCLUSION： The small molecule Akt inhibitor perifosine shows substantial anti-tumor activity in human gastric cancer cell line SGC-7901.Perifosine induces caspase-dependent apoptosis,and the key regulators include caspase-3,caspase-9 and Bcl-2.