藤梨根提取物对人胃癌MKN-45细胞增殖及凋亡的影响

摘要

目的:探讨太白山藤梨根提取物(Radix Actinidiae extractive,RAE)在体外对胃癌MKN-45细胞增殖与凋亡的影响.方法:用0.01-100μg/ml浓度范围内的RAE和胃癌MKN-45细胞共培养24、48、72 、96小时,采用MTT法检测RAE对MKN-45细胞增殖的影响;用流式细胞仪检测其对MKN-45细胞凋亡及细胞周期分布的影响;DNA琼脂糖凝胶电泳检测细胞凋亡.结果:RAE在体外能够显著抑制胃癌MKN-45细胞的增殖,且呈现浓度和时间依赖性;RAE对胃癌细胞的作用表现为周期特异性,使细胞阻滞于 G0/G1 期,进一步诱导细胞凋亡.对照组凋亡率为0.53%,1μg/ml 、10μg/ml 、100μg/ml浓度的RAE干预胃癌细胞 48 小时后均出现凋亡峰,细胞凋亡率分别为3.27%、8.97%、12.6%.DNA琼脂糖凝胶电泳检测细胞凋亡,不同浓度RAE处理的细胞样品中均可看到梯形凋亡条带,并呈现浓度依赖性.结论:RAE在体外对人胃癌 MKN-45细胞有显著的抑制细胞增殖及诱导细胞凋亡作用,其作用机制可能是使胃癌细胞周期阻滞于G0/G1期.%Objective :To investigate the role of Radix Actinidiae extractive( RAE) on anti - proliferation and pro - apoptosis for gastric cancer MKN - 45 cell line in vitro.Methods : MKN - 45 cells were exposed to different concentrations of RAE for 24h , 48h,72h and 96h respectively.The proliferation was detected by 4 - methyl - teerazolium ( MTT) , the distrihution of cell cycle and effect on apoptosis of RAE were analyzed by flow cytometry( FCM).DNA gel electrophoresis was used to demonstrate DNA fragmentation.Results : RAE significantly inhibited the proliferation of gastric cancer cells in vitro on a dose - and time dependent manner; The result of FCM made it clear that RAE showed cell cycle specificity and arrested at Go/G1.After treating 48 hours on the gastric cancer cells with RAE,all appearance apoptosis peak , the apoptosis rates were 3.27％ ,8.97％ , 12.6％ , compared to control group 0.53％ ( P ＜ 0.05 ) ; DNA agorose gel electrophoresis( AGE) analysis revealed that RAE pretreatment lead to DNA tragmentation on a dose - dependent manner.Conclusion : RAE significantly inhibited the proliferation and induced apoptosia of gastric cancer MKN -45 ceUs in vitro, which arrested cell cycle at Co/G1 .