盐霉素联合17-AAG促进人乳腺癌细胞MCF-7凋亡的机制研究

Mechanism of salinomycin combined with 17-AAG to promote apoptosis in human breast cancer cell line MCF-7

目的探讨盐霉素与17-AAG联合用药对人乳腺癌MCF-7细胞凋亡的作用及机制。方法CCK8法检测单药及联合用药对MCF-7细胞增殖的影响;用流式细胞术检测单药及联合用药对MCF-7细胞凋亡的影响;用qRT-PCR检测与凋亡相关基因的表达情况。结果细胞增殖实验显示加药后48 h药物杀伤作用更为明显,且联合用药组的细胞增殖抑制率明显高于其他对照组。流式细胞术测定结果显示空白对照组、盐霉素单药组、17-AAG单药组细胞早期凋亡率分为(4.24±0.90)%、(4.79+0.89)%、(2.78±1.10)%,而联合用药组早期凋亡率为(6.42±1.70)%,明显高于其他三组,组间比较差异均有统计学意义(P<0.05);qRT-PCR结果显示与其他三个对照组相比,联合用药组中Bcl-2的表达量显著降低,Caspase-3、Beclin-1、Bax的表达量显著增高,组间比较差异具有统计学意义(P<0.05)。结论盐霉素与17-AAG联合用药可能通过相关信号通路如PI3K/Akt/mTOR、MAPK、JNK等,调控凋亡基因Bcl-2、Beclin-1、Bax等的表达而促进MCF-7细胞凋亡。

Objective To investigate the effect and mechanism of salinomycin combined with 17-AAG on apoptosis of human breast cancer MCF-7 cells.Methods CCK8 method was used to detect the effect of single drug and combined drug on MCF-7 cell proliferation.Flow cytometry was used to detect the effect of single drug and combination drug on apoptosis of MCF-7 cells.The expression levels of apoptosis-related genes were detected by qRT-PCR.Results The cell proliferation experiment showed that the killing effect of the drug was more obvious 48 h after the addition of the drug,and the inhibition rate of cell proliferation in the combined drug group was significantly higher than that in the other control group.The results of flow cytometry showed that the apoptosis rate of blank control group,Sal monotherapy group and 17-AAG monotherapy group were divided into(4.24±0.90)%,(4.79+0.89)%and(2.78±1.10)%,while the early apoptosis rate of the combination therapy group was(6.42±1.70)%,which was significantly higher than that of the other three groups.The differences between groups were statistically significant(P<0.05).qRT-PCR results showed that compared with the control group of the other three groups,the expression of Bcl-2 was significantly decreased in the combination group,while the expression of Caspase-3,Beclin-1 and Bax was significantly increased,with statistical significance(P<0.05).Conclusion The combination of salinomycin and 17-AAG can promote the apoptosis of MCF-7 cells by regulating the expression of apoptotic genes Bcl-2,Beclin-1 and Bax through related signaling pathways such as PI3K/Akt/mTOR,MAPK and JNK.