摘要
目的探讨人参皂苷代谢产物CK对卵巢癌细胞CAOV3的增殖抑制作用,及其诱导细胞凋亡的机制。方法采用MTT比色法测定不同浓度(0、1.0、2.5、5.0、10.0、20.0、50.0μg/ml)CK对卵巢癌细胞CAOV3增殖的抑制作用。流式细胞术检测不同浓度(0、2.5、5.0μg/ml)CK诱导CAOV3细胞凋亡的情况;4′,6-二脒基-2-苯基吲哚(4′,6-diamidino-2-phenylindole,DAPI)染色观察细胞凋亡过程中形态变化;体外Caspase活力测定凋亡相关蛋白Caspase-3、Caspase-8和Caspase-9的激活情况;Western blot分析Caspase-3底物多聚(ADP-核糖)聚合酶[Poly(ADP-ribose)polymerase,PARP]的断裂情况。结果 CK可明显抑制卵巢癌细胞CAOV3的增殖,CK浓度与其对细胞的增殖抑制作用呈剂量-效应关系,IC50值为3.6μg/ml。随着CK浓度的增加,发生凋亡的细胞数增加,出现典型的凋亡形态特征的细胞逐渐增多。Caspase-3、Caspase-8和Caspase-9均被激活。随着CK浓度的增加,酶活性增加,Caspase-3底物PARP断裂的条带逐渐加深。结论 CK对卵巢癌细胞CAOV3有毒性作用,能够抑制细胞增殖,且CK浓度与这种抑制作用呈剂量-效应关系;CK抑制CAOV3细胞的增殖作用是通过促进细胞凋亡来实现的,且这种细胞凋亡涉及内源及外源型途径。
Objective T o investigate the inhibitory effect of metabolic product CK of ginsenoside on proliferation of ovarian cancer CAOV3 cells as well as its mechanism in induction of cell apoptosis. Methods The inhibitory effect of CK at various concentrations（0, 1. 0, 2. 5, 5. 0, 10. 0, 20. 0 and 50. 0 μg / ml）on proliferation of ovarian cancer CAOV3 cells was determined by MTT assay, while the apoptosis of CAOV3 cells induced with CK at various concentrations（0,2. 5 and 5. 0 μg / ml） by flow cytometry. The morphological change of cells during apoptosis was observed by 4 ′, 6-diamidino-2-phenylindole（DAPI）staining, while the activation of Caspase-3,-8 and-9 by in vitro Caspase activity assay,and the cleavage of poly（ADP-ribose） polymerase（PARP） by Western blot. Results CK showed significantly dosedependent inhibitory effect on the proliferation of CAOV3 cells, with an IC50 of 3. 6 μg / ml. The counts of apoptotic cells and the cells with typical morphological features of apoptosis increased with the increasing CK concentration. Caspase-3,-8 and-9 were activated. The activity of Caspase-3 increased with the increasing CK concentration, while the band of cleaved PARP was getting dark. Conclusion CK showed toxic effect on ovarian cancer CAOV3 cells, which inhibited the cell proliferation in a dose-dependent mode. CK inhabited the proliferation of CAOV3 cells by promoting the cell apoptosis involving endogenous and exogenous pathway.
出处
《中国生物制品学杂志》
CAS
CSCD
2015年第2期142-146,共5页
Chinese Journal of Biologicals
基金
国家自然科学基金(31240078)
2012年吉林省人民政府人才开发资助金
