桂枝茯苓丸联合顺铂对人卵巢癌SKOV-3细胞增殖与凋亡的影响

Effects of cinnamon Twig and poria pill combined with cisplatin on proliferation and apoptosis of skov-3 cells in human ovarian cancer

目的探讨桂枝茯苓丸联合顺铂对人卵巢癌SKOV-3细胞增殖、凋亡的影响。方法将48只雌性SD大鼠随机分为生理盐水(NS)组、桂枝茯苓丸组、顺铂(DDP)组、桂枝茯苓丸+DDP(联合)组;成年雌性SD大鼠经药物处理后取血制备含药血清;MTT法检测含药血清对SKOV-3细胞增殖的影响;流式细胞仪检测各组含药血清对SKOV-3细胞周期的影响;Western blot检测Bcl-2和Cyt C、Cleaved-Caspase-3蛋白表达水平。结果MTT法检测结果显示,与NS组相比,桂枝茯苓丸联合顺铂干预能显著抑制SKOV-3细胞增殖活性;流式细胞仪检测结果显示,含药血清处理SKOV-3细胞48h后,与NS组相比,DDP组、桂枝茯苓丸组、联合组G0/G1期比例上升,而S期比例下降,差异具有统计学意义(P<0.05);Western blot检测发现,与NS组相比,DDP组、桂枝茯苓丸组、联合组SKOV-3细胞内Cyt C、Cleaved-Caspase-3蛋白表达显著增加,而Bcl-2蛋白表达明显下降,差异具有统计学意义(P<0.05);与DDP组、桂枝茯苓丸组相比,联合组SKOV-3细胞内Cyt C、Cleaved-Caspase-3蛋白表达增加,而Bcl-2蛋白表达下降,差异具有统计学意义(P<0.05)。结论桂枝茯苓丸能够显著增强顺铂对SKOV-3细胞增殖抑制作用,可能是通过线粒体途径诱导其凋亡发挥作用。

Objective To investigate the effect of cinnamon Twig and Poria pill combined with cisplatin on the proliferation and apoptosis of skov-3 cells in human ovarian cancer.Methods 48 female adult SD rats were randomly divided into normal saline group,cinnamon Twig and Poria Pill group,cisplatin group,cinnamon Twig and Poria pill plus cisplatin intervention group;The cultivated skov-3 cells were randomly divided into control group,normal saline group,cinnamon Twig and Poria Pill group,cisplatin group,and cinnamon Twig and Poria pill plus cisplatin invention group,all rats were feed and applied intraperitoneal injection for 5 days continuously.The blood of rats were extracted after deep anesthetization,and serum was centrifuged.The serum containing drugs in different concentrations were applied to skov-3 cells of human ovarian cancer for 24h and 48h,In addition,the proliferation of skov-3 cells in human ovarian cancer were detected by MTT method,and the cell cycle were detected by flow cytometry,and the levels of bcl-2,Cyt C and Cleaved Caspase 3 were detected by Western blot.Results MTT assay showed that compared with normal saline group,the proliferation activity of skov-3 cells in cinnamon Twig and Poria pill plus cisplatin intervention group was significantly lower than other groups.Flow cytometry showed that after intervention in skov-3 cells for 48 hours,compared with the normal saline group,the G0/G1 phase ratio was significantly increased in the DDP group,the cinnamon Twig and Poria Pill group and the cinnamon Twig and Poria pill plus cisplatin intervention group,as opposed to the S phase ratio was obviously decreased(P<0.05).Compared with the normal saline group,western blot analysis showed that the expression levels of Cyt C and cleaved caspase-3 proteins in skov-3 cells were significantly out performed in the DDP group,the cinnamon Twig and Poria pill group and cinnamon Twig and Poria pill plus cisplatin invention group,as opposed to the expression of bcl-2 protein was dramatically decreased in the skov-3 group(P<0.05).Compared to the DDP group and the cinnamon Twig and Poria pill group,the levels of Cyt C and cleaved caspase-3 in skov-3 cells were markedly increased,as opposed to the expression of bcl-2 were markedly decreased in cinnamon Twig and Poria pill plus cisplatin intervention group(P<0.05).Conclusion Cinnamon Twig and Poria pill combined with cisplatin can dramatically enhance the inhibition of skov-3 cell proliferation and the inducement of its apoptosis by mitochondrial pathway.