Objective To investigate the effects of valproic acid (VPA),one of histone deacetylase (HDAC) inhibitors,single or combinations with paclitaxel (PTX),on the cell cycle and its mechanism of the lung cancer cell line A549 cell.Methods Experimental groups:blank control group,paclitaxel group,valproic acid group,paclitaxel combined with valproic acid.Effect of flow cytometry to detect the cell cycle ofA549 cells.The determination of differences in expression of survivin and p21 gene by fluorescence quantitative PCR method;Determination of expression of Survivin and p21 protein in cells by Western blot.Results Flow cytometric detection of VPA combined with PTX on cell cycle of lung cancer A549 cells.The four groups ofA549 cell were cultivate for 24 h,flow cytometry assay confirmed the cell cycle ofA549 cell line,the G0/G1 stage rate ofA549 in VPA and VPA combined with PTX groups are higher than other groups,and the rate of combined treatment group is the highest,the difference was statistically significant (P ＜ 0.05).Fluorescence quantitative PCR and Western blot to detect the two drugs combined effects on the expression of Survivin and p21 gene and protein in A549 lung cancer cell line.The expression of Survivin gene and protein in each treatment group decreased and combination group in lowest level.The p21 gene and protein expression increased,and the combined group was the highest (P ＜ 0.05).The difference between the groups was statistically significant.Conclusion VPA can inhibit A549 cells in G0/G1 stage.VPA combined with PTX,promote the inhibition ofA549 cell.The mechanism may be related to up-regulate the expression of p21,down-regulation of Survivin in A549 cells.%目的 研究丙戊酸钠联合紫杉醇对人肺腺癌A549细胞周期的影响,探讨其可能的相关分子机制.方法 设立空白对照组、紫杉醇组、丙戊酸钠组、丙戊酸钠联合紫杉醇组,流式细胞术检测各组A549细胞周期,荧光定量PCR及蛋白免疫印迹法测定各组A549细胞Survivin、p21基因及蛋白的表达.结果 丙戊酸钠组和丙戊酸钠联合紫杉醇组G0/G1期细胞百分比明显高于空白对照组及紫杉醇组,且丙戊酸钠联合紫杉醇组最高,差异有统计学意义(P＜0.05).药物作用24h后,与空白对照组比较,各组A549细胞Survivin基因及蛋白表达均明显降低,丙戊酸钠联合紫杉醇组表达最低(P＜ 0.05);p21基因及蛋白的表达均升高,丙戊酸钠联合紫杉醇组的表达最高(P＜0.05).结论 丙戊酸钠协同紫杉醇阻滞人肺腺癌A549细胞于G0/G1期,其机制可能与其下调A549细胞Survivin、上调p21表达有关.
展开▼
