组蛋白去乙酰化酶抑制剂调控p21^(WAF1/CIP1)启动子乙酰化水平影响乳腺癌MCF-7细胞周期

SAHA affects cell cycle of MCF-7 breast cancer cells by regulating acetylated levels of p21WAF1/CIP1promoter

目的研究组蛋白去乙酰化酶抑制剂(histone deacetylase inhibitor,HDACI)调控p21^(WAF1/CIP1)启动子乙酰化水平,调节乳腺癌MCF-7细胞周期的分子机制。方法采用实时定量PCR、Western blot和DNA-Ch IP方法测定SAHA对乳腺癌MCF-7细胞周期调控系统的影响;应用染色质免疫沉淀(chromatin immunoprecipitation,Ch IP)技术探究SAHA调控p21^(WAF1/CIP1)启动子乙酰化水平的情况。结果 SAHA明显影响乳腺癌细胞周期相关调控因子的表达;在针对p21^(WAF1/CIP1)基因功能的筛查中发现,SAHA可明显诱导p21^(WAF1/CIP1)mRNA和蛋白的表达,并且可调节p21^(WAF1/CIP1)启动子乙酰化水平。结论 SAHA通过影响p21^(WAF1/CIP1)启动子乙酰化程度调节乳腺癌MCF-7细胞周期的进程。

Aim To study the regulation mechanisms of deacetylase inhibitor SAHA in p21（WAF1/CIP1） promoter acetylation in breast cancer MCF-7 cells. Methods We used quantitative real-time PCR,Western blot and DNA-Ch IP to determine the effects on the regulation of cell cycle with SAHA treatment in MCF-7 cells. By DNA-Ch IP, we assessed the acetylation level of p21（WAF1/CIP1） promoter. Results SAHA significantly affected the expression of cell cycle-related factors,and induced the mRNA and protein expression of p21（WAF1/CIP1）. SAHA could adjust the acetylation level of p21（WAF1/CIP1） promoter. Conclusion SAHA regulates the cell cycle progression by adjusting the acetylation level of p21（WAF1/CIP1） promoter in MCF-7 cells.