青蒿琥酯对人结肠癌细胞HCT-8细胞凋亡和细胞周期的影响

目的:研究青蒿琥酯对人结肠癌HCT-8细胞凋亡和细胞周期的影响.方法:实验分为10,20,30 μmol·L-1青蒿琥酯处理组,阴性对照组和空白对照组.采用透射电镜和流式细胞仪检测青蒿琥酯对HCT-8细胞的凋亡诱导效果；采用流式细胞仪分析青蒿琥酯对HCT-8细胞周期的影响；采用Western blot印迹法检测青蒿琥酯对细胞凋亡相关蛋白Bax和Bcl-2表达的影响.结果:经青蒿琥酯处理后,电镜下可见HCT-8细胞膜皱缩、染色质凝聚、核碎裂、凋亡小体形成.10,20,30 μmol·L-1青蒿琥酯处理组凋亡率分别为17.1％±3.8％,29.5％±5.1％,41.4％±5.8％,显著高于空白对照组5.1％±1.4％.P＜0.05.在20 μmol· L-1青蒿琥酯处理组,HCT-8细胞G0/G1期所占比例随着药物作用时间延长而增加,S期与G2/M期细胞所占比例则下降.10,20,30 μmol·L -1青蒿琥酯处理组Bax蛋白表达水平分别为0.20±0.03,0.40±0.05和0.50±0.08显著高于空白对照组(0.06 ±0.02),P＜0.05；Bcl-2蛋白表达水平未见明显变化,Bcl-2/Bax呈下降趋势.结论:青蒿琥酯可以抑制人结肠癌细胞HCT-8增殖,诱导细胞凋亡,其诱导凋亡作用机制可能与其阻滞结肠癌细胞周期和上调促癌基因Bax表达有关.

Effect of Artesunate on Cell Apoptosis and Cell Cycle of Human Colon Cancer Cell HCT-8

Objective:To investigate the effect of artesunate on apoptosis and cell cycle of human colon cancer HCT-8 cells.Method: The experimental groups were negative control group,blank control group,10,20 and 30 μmol·L-1 artesunate-treated groups.transmission electron microscope(TEM) and flow cytometry(FCM) were used to analyze the apoptosis of the treated HCT-8 cells by artesunate.FCM was used to analyse cell cycle of the treated HCT-8 cells by artesunate.The levels of Bax and Bcl-2 involved in the different treated HCT-8 cells were detected by Western blot.Result: After treatment of artesunate,it had observed by TEM that the cell shape changed with cytomembrane shrink,karyopycnosis,nuclear fragmentation,and the formation of apoptotic bodies.The rates of apoptosis of 10,20,30 μmol·L-1 artesunate-treated groups were 17.1%±3.8%,29.5%±5.1%,41.4%±5.8%,which was significantly higher than the apoptosis rare of blank control group(5.1%±1.4%,P<0.05).In 20 μmol·L-1 artesunate-treated group,G0/G1 cell proportion was increased with the prolonged drug effects,and S and G2/M cell proportion was decreased.Bax protein expression levels of 10,20,30 μmol·L-1 artesunate-treated groups were 0.20±0.03,0.40±0.05,0.50±0.08,which was significantly higher than that in blank control group(0.06±0.02,P<0.05).The difference of Bcl-2 protein expression levels in the above-mentioned groups was not statistically significant.The protein expression ratio of Bcl-2/Bax showed downward trend.Conclusion: Artesunate can inhibit human colon cancer HCT-8 cell growth and induce apoptosis of HCT-8 cell lines by blocking cell cycle and regulating the expression of Bax genes.