红花黄色素对人肝癌细胞凋亡及相关蛋白表达的影响

Effect of Safflower Yellow on Apoptosis and Expression of Related Proteins in Human Hepatocellular Carcinoma Cells

目的:探讨红花黄色素(safflor yellow,SY)对MHCC-97H细胞凋亡及相关蛋白表达的影响。方法:选用MHCC-97H细胞体外培养至对数生长期,加入不同浓度SY(0、0.03125、0.0625、0.125、0.25、0.5 mg/L)进行体外培养,并设顺铂阳性对照组(终浓度0.5μg/ml),分别干预24 h、48 h及72 h后,采用四甲基偶氮唑蓝(MTT)法观察SY对细胞增殖的抑制作用,流式细胞仪观察细胞周期及凋亡率的变化,Western–blotting检测Bax、Bcl-2蛋白表达情况。结果:MTT显示SY对人肝癌细胞MHCC-97H增殖有一定的抑制作用,与SY浓度及作用时间有相关性;流式细胞术显示,细胞凋亡率随SY浓度增加有上升趋势,细胞周期无明显变化;Western-blotting显示,随着SY浓度的增加,Bax表达增加,Bcl-2表达减少,而Bcl-2/bax比值下降。结论:SY对MHCC-97H细胞的生长有一定的抑制作用,但对细胞周期无明显阻滞作用,其可诱导该细胞凋亡,下调Bcl-2/Bax比值,可能为其促凋亡机制之一。

Objective： To investigate the effect of safflower yellow（ SY） on apoptosis and related protein expression in MHCC-97 H cells. Methods： MHCC-97 H cells cultured in vitro in logrithmic growth phase wered selected. Then different concentrations（ 0,0. 03125,0. 0625,0. 125,0. 25,0. 5 mg / L） of SY were added to cell culture medium in matched experimental groups,and positive control group to Cisplatin（ at concentration of 0. 5μg / ml）. Respectively intervene of 24 h,48 h and 72 h,Inhibition on cell proliferation of experimental groups was evaluated by four methyl thiazolyl tetrazolium（ MTT） method. The changes of cycle and apoptosis rate were detected with flow cytometry. Expression of Bax and Bcl-2protein in all experimental groups was detected by Western blotting method. Results： MTT results show that,SY has a certain inhibitory effect on the proliferation of human hepatoma MHCC-97 H cells,is correlated with SY concentration and action time. Flow cytometry analysis show that,the apoptosis rate decreased with increasing SY concentration has an upward trend, no significant changes in cell cycle. Western-blotting results show that, with the increase of SY concentration,the expression of Bax was increased,Bcl-2 expression decreased,and the decrease of the ratio of Bcl-2 /bax. Conclusion： A certain degree of inhibition of SY on the growth of MHCC-97 H cells and induce its apoptosis,no obvious blocking effect on cell cycle,by decreasing the ratio of Bcl-2 / Bax may be one of mechanisms of promoting apoptosis.