Wortmannin与ABT-737诱导卵巢癌细胞凋亡的协同作用研究

目的:探讨磷酸肌醇3-激酶(PI3K)抑制剂wortmannin单用以及与Bcl-2蛋白抑制剂ABT-737联合应用对人卵巢癌细胞系存活率和凋亡率的影响。方法:①CCK-8法分别测定Wortmannin、ABT-737单独以及联合应用对人卵巢癌细胞ES-2和SKVO-3生长的抑制率,并计算其IC50。②Annexin-V/PI法流式细胞仪检测细胞凋亡率以及对细胞周期的影响,Western blot法检测相关蛋白的表达水平,分别测定Wortmannin、ABT-737单独以及联合应用对人卵巢癌细胞ES-2和SKVO3凋亡的影响。结果:①Wortmannin和ABT-737单独作用对ES-2细胞株的IC50分别为14.12μM和31.31μM,两药联合应用的IC50为8.15μM;Wortmannin和ABT-737单独作用对SKVO-3细胞的IC50分别为16.72μM和37.56μM,联合用药的IC50为9.25μM。②0.2μM的Wortmannin作用48 h后ES-2和SKVO-3细胞的凋亡率分别为9.9%和8.7%,5μM的ABT-737作用48 h后ES-2和SKVO-3细胞凋亡率分别为10.2%和9.1%,两者联合应用对两种细胞的凋亡率分别为45.0%和53.0%,组间比较差异有统计学意义(P<0.01)。③Wortmannin和ABT-737联合应用可以明显的增加cleaved-caspase-3、细胞色素C以cleaved-PARP的表达量,Wortmannin和ABT-737联合应用可以明显的对人卵巢癌ES-2和SKVO-3细胞周期阻滞在G0/G1期。结论:Wortmannin和ABT-737能够通过线粒体途径抑制卵巢癌细胞株的生长并诱导其凋亡,且Wortmannin和ABT-737联合应用具有协同作用。

Synergy effect of Wortmannin and ABT-737 on the induction of apoptosis in ovarian cancer cells

Objective:To investigate the survival rate and apoptotic rate of human ovarian cancer cells induction by both the phosphatidylinosito1 3-kinase(PI3K) inhibitor Wortmannin and the Bcl-2 inhibitor ABT-737 and their synergistic effect.Methods:①CCK8 assay was used to examined inhibiting rate of human ovarian cancer cells of ES-2 and SKOV-3.After Wortmannin,ABT-737 single and combination application and IC50was calculated.②Annexin-V/PI assay was applied to examine the cell apoptosis rate and cell cycle.Additionally,Western blot was introduced to evaluate the expression levels of cleaved-caspase-3 and cleaved-PARP.Moreover,cytochrome C in cytoplasm was detected.Results:①IC50of Wortmannin and ABT-737 single and combination application for ES-2 was 14.12,31.31,8.15 μmol/L respectively.IC50of Wortmannin and ABT-737 single and combination application for SKOV-3 was 16.72,37.56,9.25 μmol/L respectively.②Apoptosis rate of ES-2 and SKOV3 was 9.9% and 8.7% by 0.2 μmol/L Wortmannin single application for 48 h respectively,10.2% and 9.1% by 5 μmol/L ABT-737 single application for 48 h respectively,and 45.0% and 53.0% by Wortmannin and ABT-737 combination application for 48 h respectively.There were all statistical differences(P<0.01).③Expressions of cleaved-caspase-3,cytochrome C and cleaved-PARP were obviously increased and cell cycles were blocked at G0/G1 phase by Wortmannin and ABT-737 combination application.Conclusion:Wortmannin and ABT-737 could inhibit the growth and induce apoptosis of ovarian cancer cells through the mitochondrial pathway and Wortmannin and ABT-737 have the synergetic effect.