SIRT1在宫颈癌细胞耐药中的作用及其机制研究

摘要目的：探讨沉默交配型信息调节因子2同源蛋白1（silentmatingtypeinformationregulation2homologyl，SIRTl）在宫颈癌化疗耐药中的作用及其机制。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，westernblotting检测MMC对Hela和Hela／MMC细胞内SIRTl蛋白表达的影响；MTT法检测MMC及Nicotinamide对Hela和Hela／MMC细胞增殖的影响；AnnexinV-PI试验检测Hela／MMC细胞凋亡的亡的情况；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／MMC细胞中SIRTl的表达显著高于Hela细胞（P〈0．05），MMC处理的Hela／MMC细胞中SIRTl的表达显著高于未经MMC处理（P〈0．05）。Nicotinamide对Hela和Hela／MMC细胞具有相似的生长抑制作用，Nicotinamide可使MMC诱导的Hela／MMC细胞凋亡增加，同时降低细胞内P-gP的mRNA表达（P〈0．05）。结论：SIRTl表达下调能显著减轻Hela／MMC细胞对MMC的耐药性，其作用可能与P—gp有关。

Role of SIRT 1 in Drug Resistance of Cervical Cancer Cells

Objective： To discuss the role and mechanism of silent mating type information regulation 2 homology 1 （SIRT1） 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 SIRT1 protein expression was detected by Western blotting; MTT assay was used to detect the proliferative effect of Nicotinamide and MMC on Hela and Hela/MMC cells. Apoptotic cells were detected with Annexin V staining; mRNA of drug resistant-associated protein P-gp was measured by RT-PCR. Results： The expression of SIRT1 protein in Hela/MMC was significantly higher than that in Hela cells normally （P〈0.05） and the expression of SIRT1 protein in Hela/MMC MMC-treatment group was significantly higher than that in control （P〈0.05）; SIRT1 inhibitors Nicotinamide exhibited the similar growth-inhibitory effects on Hela and Hela/MMC, which can induce the increase of Hela/MMC cell apoptosis and also reduce the mRNA level of P-gp （P〈0.05）. Conclusion： The downregulation of SIRT1 can significantly mitigate MMC- resistance of MMC-Hela, which may be achieved by regulated P-gp expression.