摘要
目的探讨黄芪多糖协同顺铂诱导宫颈癌siha细胞凋亡的作用机制。方法 siha细胞分为4组:对照组、黄芪多糖组、顺铂组、黄芪多糖联合顺铂组,采用四甲基偶氮唑盐比色法(MTT比色法)观察各组siha细胞的增殖抑制作用;Annexin V-FITC/PI双染法检测各组细胞的凋亡率,蛋白印迹法(Western blot法)检测各组细胞凋亡相关蛋caspase3、caspase7、caspase9和Bcl-2的表达情况。结果 MTT结果显示,各用药组siha细胞增殖于24、48、72 h均可被抑制,与对照组相比,黄芪多糖组和顺铂组对siha细胞有明显的增殖抑制作用(P<0.05),黄芪多糖联合顺铂组的抑制作用更加显著(P<0.01);与黄芪多糖组和顺铂组相比,联合用药对siha细胞有显著的抑制作用(P<0.05)。流式细胞仪检测结果显示:与对照组相比,黄芪多糖组和顺铂组的凋亡率明显增加(P<0.05),黄芪多糖联合顺铂组的凋亡率显著增加(P<0.01)。蛋白印迹法(Western blot法)检测结果显示:与对照组相比,黄芪多糖组、顺铂组中caspase3、caspase7、caspase9蛋白表达水平明显升高(P<0.05),Bcl-2蛋白表达水平明显下降(P<0.05);黄芪多糖联合顺铂组的上述变化更加显著(P<0.01)。结论黄芪多糖可以协同顺铂诱导宫颈癌siha细胞凋亡,这一作用的机制可能是通过抑制Bcl-2蛋白的表达,从而增加caspase9的募集,启动caspase级联反应,进一步激活下游的caspase3、caspase7,促进细胞凋亡。
Objective To investigate the mechanism of Astragalus Polysaccharides combined with cisplatin on the apoptosis of cervical cancer cell line siha. Methods Siha cells were divided into four groups： control group,Astragalus Polysaccharide group,cisplatin group and Astragalus Polysaccharide combined with cisplatin group. MTT colorimetric method was used to observe the proliferation of siha cells. The expression of caspase 3,caspase 7,caspase 9 and Bcl-2 in the cells were assessed by Western blot. Results MTT results showed that siha cells were proliferated at 24,48 and 72 h,and the proliferation of siha cells was significantly inhibited by Astragalus Polysaccharide and cisplatin compared with the control group（ P 〈 0. 05,P 〈 0. 01）. Compared with Astragalus Polysaccharide and cisplatin alone group,the combination therapy had significant inhibitory effect on siha cells（ P 〈 0. 05）. The apoptotic rates of Astragalus Polysaccharide group and cisplatin group were significantly higher than that of control group（ P 〈 0. 05）. The apoptotic rate of Astragalus Polysaccharide combined with cisplatin was significantly increased（ P 〈 0. 01） The expression of caspase 3,caspase 7 and caspase 9 protein in Astragalus Polysaccharide group and cisplatin group were significantly higher than those in control group（ P 〈 0. 05）,and the expression level of Bcl-2 protein was significantly reduced（ P 〈 0. 05）. The changes of Astragalus Polysaccharide combined with cisplatin were more significant（ P 〈 0. 01）. Conclusion Astragalus Polysaccharide can induce apoptosis of cervical cancer cell line siha cells in combination with cisplatin. The mechanism of this action may be through the inhibition of Bcl-2 protein expression,thus increasing the caspase9 recruitment,initiation of caspase cascade reaction,further activation of downstream caspase 3,caspase 7 and cell apoptosis.
作者
翟秋丽
胡向丹
肖静
余冬青
ZHAI Qiu - li;HU Xiang - dan;XIAO Jing;YU Dong - qing(Second Clinical Medical College, Guangzhou University of Traditional Chinese Medicine, Guangzhou 510006, China)
出处
《广东医学》
CAS
2018年第9期1327-1331,共5页
Guangdong Medical Journal
基金
国家自然科学基金资助项目(编号:81403422)
关键词
子宫颈癌
SIHA细胞
黄芪多糖
顺铂
协同效应
cervical cancer
siha cell
Astragalus Polysaccharide
cisplatin
synergistic effect
