伏立诺他与紫杉醇增强ROS水平介导Hela细胞凋亡机制研究

Study on the mechanism of Hela cell apoptosis mediated by vorinolta and paclitaxel enhancing ROS level

目的:研究组蛋白去乙酰化酶抑制剂-伏立诺他(SAHA)联合紫杉醇促进宫颈癌Hela细胞凋亡的机制。方法:采用流式细胞术检测SAHA、紫杉醇及SAHA联合紫杉醇对Hela细胞活性氧(ROS)含量和细胞凋亡率的影响,Western blot法检测Hela细胞中凋亡相关蛋白caspase-3和caspase-9表达,免疫荧光法观察不同药物处理后Hela细胞微管结构的变化。结果:SAHA+紫杉醇组Hela细胞中ROS水平高于SAHA组和紫杉醇组,SAHA+紫杉醇组与紫杉醇组有明显统计学差异(P<0.05)。SAHA+紫杉醇组Hela细胞的凋亡率明显高于对照组、紫杉醇组、SAHA组(P<0.01,P<0.05,P<0.01)。SAHA+紫杉醇组的caspase-3和caspase-9蛋白表达高于紫杉醇组和SAHA组(P<0.05)。SAHA和紫杉醇联合作用改变Hela细胞的微管结构,使微管短小纤细,向细胞核靠拢。结论:组蛋白去乙酰化酶抑制剂(HDACi)-伏立诺他(SAHA)与紫杉醇联合作用可显著促进细胞凋亡,其机制可能是通过影响Hela细胞氧化应激机制,促进Hela细胞凋亡。

Objective:To investigated the apoptotic mechanism of histone deacetylase inhibitor volinota(SAHA)combined with paclitaxel in cervical cancer Hela cells.Methods:Flow cytometry was used to detect the effects of SAHA,paclitaxel and SAHA combined with paclitaxel on the content of reactive oxygen species(ROS)and apoptosis rate of Hela cells.Western blot was used to detect the expression of caspase-3 and caspase-9 in Hela cells.Immunofluorescence was used to observe the changes of microtubule structure of Hela cells treated with different drugs.Results:ROS levels in Hela cells of SAHA combined with paclitaxel group were significantly higher than those of SAHA and paclitaxel group,and there was significant difference(P<0.05).The apoptosis rate of Hela cells in the SAHA paclitaxel group was significantly higher than that in the SAHA and paclitaxel groups(P<0.01,P<0.05).The expression of caspase-3 and caspase-9 in SAHA combined with paclitaxel group was higher than that in paclitaxel group and SAHA group(P<0.05).The microtubules of Hela cells treated with SAHA and paclitaxel for 24 hours were short and slender,and tended to approach the nucleus.Conclusion:The combination of HDACi(SAHA)and paclitaxel can significantly promote apoptosis,which may be related to their involvement in mitochondrial-derived apoptotic mechanisms such as oxidative stress in Hela cells.