水飞蓟宾与阿霉素联合作用对胃癌细胞SGC-7901凋亡的影响及其机制

Effect of silibinin combined with doxorubicin on apoptosis of gastric cancer SGC-7901 cells and relevant mechanism

目的探讨水飞蓟宾(silibinin)与阿霉素(doxorubicin)联合作用对胃癌细胞SGC-7901凋亡的影响及其机制。方法用不同浓度的水飞蓟宾、阿霉素单独或联合作用SGC-7901细胞,采用MTT法检测细胞增殖的活力;流式细胞术检测细胞周期和细胞凋亡的变化;Western blot法检测细胞周期调控蛋白Cdk1、Cyclin B1和细胞凋亡调控蛋白pro-caspase 3、pro-caspase 8、pro-caspase 9、PARP、Fas、Fas L表达量的变化及Bcl-2/Bax的比例。结果 0.01μmol/L阿霉素联合水飞蓟宾(25、50、100、200μmol/L)作用SGC-7901细胞后,产生了明显的协同作用,联合作用指数(combined index,CI)分别为0.634、0.418、0.224、0.472,有效提高了细胞的抑制率,同时降低了Cdk1和Cyclin B1蛋白的表达量,引起细胞G2/M期阻滞;Fas和Fas L的表达量升高,pro-caspase 3/8/9的表达量降低,PARP被剪切,Bcl-2/Bax的比例降低,促使SGC-7901细胞凋亡。结论水飞蓟宾与阿霉素联合作用SGC-7901细胞后,水飞蓟宾能有效降低阿霉素的使用浓度,并提升其诱导凋亡的效果。

Objective To investigate the effect of silibinin combined with doxorubicin on the apoptosis of gastric cancer SGC-7901 cells and the relevant mechanism. Methods SGC-7901 cells were treated with silibinin and doxorubicin at various concentrations, alone or in combination, and determined for proliferation activity by MTT method, for cell cycle and apoptosis by flow cytometry, and for expression levels of cell cycle regulatory proteins Cdk1 and Cyclin B1, cell apoptosis proteins pro-caspase 3, pro-caspase 8, pro-caspase 9, PARP, Fas and Fas L as well as ratio of Bcl-2 / Bax, were determined by Western blot. Results Doxorubicin at a concentration of 0. 01 μmol / L in combination with silibinin at concentrations of 25, 50, 100 and 200 μmol / L showed significantly synergistic effect on SGC-7901 cells, with combined indexes（CIs）of 0. 634, 0. 418, 0. 224 and 0. 472 respectively, which increased the inhibitory rates to cells, decreased the expression levels of Cdk 1 and Cyclin B1, resulting in the cell arrest at G2/ M phase. The expression levels of Fas and Fas L increased, while those of pro-caspase 3 / 8 / 9 decreased, PARP was cleaved, the ratio of Bcl-2 / Bax decreased, and the apoptosis of SGC-7901 was induced. Conclusion Silibinin in combaintion with doxorubicin effectively decreased the concentration of doxorubicin, and promoted the induction of apoptosis.