丙戊酸钠对Kasumi-1细胞细胞周期的阻滞作用及其机制探讨

目的 研究组蛋白脱乙酰化酶(HDAC)抑制剂丙戊酸钠(VPA)对白血病细胞系Kasnmi-1细胞周期的影响并探讨其分子机制.方法 不同浓度VPA处理Kasumi-1细胞不同时间,碘化丙锭染色流式细胞术分析细胞周期的变化.RT-PCR及Western blot方法 分析细胞周期蛋白D1(cyclin D1)及周期蛋白依赖性激酶抑制因子p21 WAF1/CIP mRNA或蛋白水平变化.结果 ①VPA抑制Kasumi-1细胞的细胞周期,使其阻滞于G0/G1期.3 mmol/L VPA处理3 d,G0/G1期细胞由(50.7±2.8)%上升至(80.7±1.7)%.②以3 mmol/LVPA处理Kasumi-1细胞不同时间,与对照组比较,3 mmol/L VPA组cyclin D1 mRNA表达水平下调;以不同浓度VPA处理Kasumi-1细胞3 d,结果 发现随着VPA药物浓度增加cyclin D1 mRNA表达水平降低,呈现剂量依赖性.③VPA诱导p21WAF1/CIP mRNA表达明显增加,呈现时间及剂量依赖性.不同浓度VPA处理细胞3 d时,随着用药浓度的增加,p21WF1/CIP蛋白相对表达水平增加,3 mmol/L VPA处理细胞2 d与0 d比较,p21 WAF1/CIP蛋白相对表达水平明显增加(2.498±0.240对0).结论 VPA可以通过对cyclin D1及周期蛋白依赖性激酶抑制因子p21WAF1/CIP调节使Kasumi-1细胞周期阻滞于G0/G1期.

Inhibitory effect of valproic acid on cell cycle of Kasumi-1 cell line and its mechanism

Objective To investigate the influence of valproie acid (VPA),a histone deacetylase (HDAC) inhibitor,on cell cycle of Kasumi-1 cells,and explore its molecular mechanism.Methods Kasumi-1 ceils were treated with VPA at different concentration and different time cell cycle changes were analyzed by flow cytometry.Cyclin D1 and p21 WAF1/CIP gene,a eyclin-dependent kinase inhibitor,were determined by semi-quantitative RT-PCR and Western blot.Results ① VPA blocked the Kasumi-1 cells in G0/G1 phase.②Compared with control group,3 mmol/L VPA treated Kasumi-1 cells for different times caused their mRNA expression of cyclin D1 decreased.Treated with VPA at different concentration for 3 days,the mRNA expression decreased in a dose-dependent manner.③ VPA induced p21 WAF1/CIP mRNA expression increased in a time- and dose-dependent manner.The p21 WAF1/CIP protein increased with increasing concentration of VPA at day 3.The p21WAF1/CIP protein significantly increased with 3 mmol/L VPA treatment for 2 d (2.498±0.240).Conclusion VPA can arrest Kasumi-1 cell in G0/G1 phase through the regulation of eyclin DI and p21 WAF1/CIP.