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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。
Objective: To investigate the effects of PI3K inhibitor wortmannin alone and in combination with Bcl-2 protein inhibitor ABT-737 on the survival rate and apoptosis rate of human ovarian cancer cell lines. Methods: ① The inhibitory rates of Wortmannin, ABT-737 alone and in combination on the growth of human ovarian cancer cells ES-2 and SKVO-3 were determined by CCK-8 assay and their IC50 values were calculated. The apoptosis rate and cell cycle were detected by Annexin-V / PI flow cytometry. Western blot was used to detect the expression of related proteins. Wortmannin and ABT-737 alone and in combination were used to detect the expression of ES- 2 and SKVO3 apoptosis. Results: ①Wortmannin and ABT-737 alone had the IC50 of 14.12μM and 31.31μM for ES-2 cell line, respectively. The IC50 of combination of the two drugs was 8.15μM. The IC50 of Wortmannin and ABT-737 alone on SKOVO-3 cells were 16.72 μM and 37.56 μM, and the IC50 for the combination was 9.25 μM. ② The apoptotic rates of ES-2 and SKVO-3 cells after treated with 0.2μM Wortmannin for 48 h were 9.9% and 8.7%, respectively. The apoptotic rates of ES-2 and SKOVO-3 cells treated with 5μM ABT-737 for 48h were Were 10.2% and 9.1% respectively. The apoptosis rate of the two cell lines was 45.0% and 53.0%, respectively. The difference between the two groups was statistically significant (P <0.01). The combined application of Wortmannin and ABT-737 can obviously increase the expression of cleaved-caspase-3 and cleaved-PARP, and the combined application of Wortmannin and ABT-737 can significantly inhibit the expression of cleaved-caspase-3 and cleaved-PARP in human ovarian cancer ES-2 and SKVO-3 cells Cycle arrest in G0 / G1 phase. Conclusion: Wortmannin and ABT-737 can inhibit the growth and induce apoptosis of ovarian cancer cell lines through mitochondrial pathway, and the combination of Wortmannin and ABT-737 have synergistic effect.