蟾毒灵通过调控Livin蛋白诱导卵巢癌细胞凋亡的实验研究

Bufalin induced ovarian cancer cell apoptosis by regulation of Livin

目的:观察蟾毒灵对卵巢癌OVCAR-3的细胞活力和凋亡的影响,并进一步探究Livin蛋白在其中的作用。方法:向OVCAR-3细胞培养基中加入不同浓度梯度的蟾毒灵(0、0.1、1、10、100、500nmol/L),利用CCK-8法检测不同剂量的蟾毒灵对OVCAR-3细胞活力的影响;通过流式细胞计数法检测加入蟾毒灵后OVCAR-3细胞的凋亡情况;利用Western blot法检测Livin蛋白及Caspase-3的表达水平。结果:不同浓度的蟾毒灵作用24、48h后能显著抑制OVCAR-3细胞的活力,且呈剂量依赖效应(P<0.05),其中48h的50%的抑制浓度(IC50)为(93.2±3.2)nmol/L;OVCAR-3细胞加入蟾毒灵后能显著增加细胞凋亡的水平(P<0.05);蟾毒灵作用OVCAR-3细胞后,Livin蛋白水平降低,而Caspase-3的剪切水平升高(P<0.05)。结论:蟾毒灵可以抑制卵巢癌细胞的细胞活力,促进癌细胞的凋亡增加,而这一过程可能受到Livin蛋白的调控。

Objective:To investigate the role of bufalin on the viability and apoptosis of ovarian cancer(OVCAR-3)cells,and further examine whether Livin was involved.Methods:Different dosages(0,0.1,1,10,100,500 nmol/L)of bufalin were added to the medium of OVCAR-3 cells,and CCK-8 assay was used to examine the effect of cell viability.Flow cytometry assay was used to detect cell apoptosis in response to bufalin.Western blot was used to detected the protein expression of Livin and Caspase-3.Results:Bufalin could significantly inhibit cell viability of OVCAR-3 cell in a concentration-dependent way(P<0.05),and half maximal inhibitory concentration(IC50)of 48 h was(93.2±3.2)nmol/L.Treatment with bufalin markedly increased the percentage of cell apoptosis in OVCAR-3 cells(P<0.05).The protein expression of cleaved Caspase-3 was increased,and Livin was decreased after treatment with bufalin(P<0.05).Conclusion:Bufalin can reduce cell viability and increase apoptosis in OVCAR-3 cells,and the anti-cancer effect of bufalin may be mediated by Livin.