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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 To investigate the inhibitory effect of ginsenoside metabolite CK on the proliferation of ovarian cancer cell line CAOV3 and its mechanism of apoptosis. Methods MTT assay was used to determine the inhibitory effects of different concentrations of CK (0, 1.0, 2.5, 5.0, 10.0, 20.0 and 50.0 μg / ml) on the proliferation of ovarian cancer cells CAOV3. Flow cytometry was used to detect the apoptosis of CAOV3 cells induced by different concentrations of CK (0, 2.5, 5.0μg / ml); 4 ’, 6-diamidino-2 -phenylindole (DAPI) staining. The activation of Caspase-3, Caspase-8 and Caspase-9 were detected by Caspase activity in vitro. The protein level of Caspase-3 was detected by Western blot - ribose) polymerase [Poly (ADP-ribose) polymerase, PARP]. Results CK could significantly inhibit the proliferation of ovarian cancer cells CAOV3. The concentration of CK was dose-dependent with its inhibitory effect on cell proliferation with IC50 value of 3.6μg / ml. With the increase of CK concentration, the number of apoptotic cells increased and the number of typical apoptotic cells increased gradually. Caspase-3, Caspase-8 and Caspase-9 are activated. With the increase of CK concentration, the enzyme activity increased, and the band of cleaved PARP of Caspase-3 substrate gradually deepened. CONCLUSION: CK has a toxic effect on CAOV3 ovarian cancer cells and inhibits cell proliferation. The CK concentration has a dose-effect relationship with this inhibitory effect. CK inhibits the proliferation of CAOV3 cells through the promotion of apoptosis. Apoptosis involves both endogenous and exogenous pathways.