Hsp90靶向抑制多肽P7与多西他赛联用对肺腺癌细胞系A549的协同抗肿瘤作用

Synergistic antitumor effect of Hsp90 targeting-inhibitory peptide P7 combined with docetaxel in lung adenocarcinoma cell line A549

目的研究热休克蛋白90(Hsp90)双功能靶向抑制多肽LPLTPLP(P7)与临床化疗药物多西他赛(DTX)联用对肺腺癌细胞系A549的影响。方法以DTX与P7单独或联合使用对A549细胞进行处理,48 h后,用CCK-8法评价细胞活性;通过流式细胞计量术检测DTX与P7单独或联合使用后的A549细胞凋亡率;用蛋白免疫印迹技术检测DTX与P7单独或联合使用对A549细胞凋亡以及耐药相关蛋白表达的影响。结果DTX与P7对细胞存活具有协同抑制作用;联合用药组细胞凋亡率为25.8%,显著高于DTX组(P<0.05);联合用药组凋亡相关蛋白Bcl-2、PARP显著下调(P<0.05);cleaved-PARP显著上调(P<0.05);耐药蛋白主要穹窿蛋白(MVP)显著下调(P<0.05)。结论DTX协同靶向性多肽P7能显著提高A549细胞的凋亡率,其机制可能与降低凋亡抑制蛋白Bcl-2表达水平、失活DNA修复酶PARP及降低耐药蛋白MVP表达有关。

Objective To investigate the effects of combination of Hsp90 bifunctional targeting-inhibitory peptide LPLTPLP(P7)with chemotherapy drug docetaxel(DTX)on lung adenocarcinoma cell line A549.MethodsDTX and peptide P7 alone or combined were incubated with A549 cell for 48 hours,thereafter the effect of the drug on cell viability was evaluated using the CCK-8 method.Flow cytometry was carried out to quantify apoptotic cells.The expression of apoptotic and drug resistant protein were assessed by Western blot analysis.ResultsDTX and P7 exhibited a synergistic effect on cell survival.The apoptosis rate of the combined drug group was 25.8%,which was significantly higher than that of DTX group(P<0.05).Apoptosis-related proteins Bcl-2 and PARP were significantly down-regulated(P<0.05),and cleaved-PARP protein was significantly up-regulated(P<0.05).Meanwhile,drug-resistant protein major vault protein(MVP)was significantly down-regulated(P<0.05).Conclusions DTX ombined with targeted peptide P7 can significantly increase the apoptosis rate of A549 cells,which may be related to expression level of Bcl-2,inactivation of DNA repair enzyme PARP and down-regulation of multidrug-resistant protein MVP.