CDK5逆转Sirt1在宫颈癌细胞耐药中的作用机制

CDK5 reverses the multidrug- resistant mechanism of Sirt1 in cervical cancer cells

目的:探讨CDK5逆转Sirt1在宫颈癌化疗耐药中的作用机制。方法:体外培养人宫颈癌Hela细胞系和宫颈癌Hela/MMC耐药细胞亚系,Western blotting检测MMC对Hela和Hela/MMC细胞内P-Sirt1蛋白表达的影响;MTT法检测Hela细胞存活率;通过加入CDK5抑制剂Roscovitine来检测CDK5使Sirt1磷酸化的作用;RT-PCR方法检测耐药相关蛋白P-gp的mRNA表达情况。结果:正常情况下,Hela细胞中P-Sirt1的表达显著高于Hela/MMC细胞(P<0.05),MMC处理的Hela细胞中P-Sirt1的表达显著高于未经MMC处理组(P<0.05)。超表达P-Sirt1会导致细胞存活率显著下降(P<0.05)。CDK5抑制剂Roscovitine可以使Hela细胞的存活率增加,耐药性相关蛋白P-gp的mRNA表达上调(P<0.05)。结论:CDK5可以使Sirt1磷酸化,增加了Hela细胞对MMC的敏感性。

Objective：To discuss the mechanism of cyclin-dependent kinase 5 （CDK5） in cervical cancer drug resistance.Methods：Human cervical cancer Hela cell lines and cervical cancer Hela/MMC drug-resistant cell sublines were cultivated in vitro.The effects of MMC on P-Sirt1 protein expression was detected by Western blotting.MTT assay was used to detect the Hela cells viability.The impact of CDK5 on Sirt1 was tested through the addition of CDK5 inhibitor Roscovitine.mRNA of drug resistant-associated protein P-gp was measured by RT-PCR.Results：The expression of P-Sirt1 protein in Hela was significantly higher than Hela/MMC cells normally （P 〈 0.05） and the expression of Sirt1 protein in Hela MMC-treatment group was significantly higher than control（P 〈 0.05）.Overexpression of P-Sirt1 leads to the significant reduction of cell viability（P 〈 0.05）.Inhibitor Roscovitine can increase cell survival and upregulate the expression of P-gp mRNA （P 〈 0.05）.Conclusion：CDK5 phosphorylate Sirt1 increase sensitivity to MMC in Hela cells.