姜黄素抑制HepG2细胞HDAC1活性及促进P21^(WAF1/CIP1)表达的研究  

Inhibition of curcumin on histone deacetylase and expression promotion of P21^(WAF1/CIP1) in HepG2 cells

作　　者：吕必华[1] 张玲[2] 朱长才[2] 刘静[3] 

机构地区：[1]武汉大学中南医院药学部,湖北武汉430071 [2]武汉科技大学医学院预防医学系,湖北武汉430065 [3]武汉大学药学院,湖北武汉430072

出　　处：《中国中药杂志》2007年第19期2051-2055,共5页China Journal of Chinese Materia Medica

摘　　要：目的:研究姜黄素(curcumin,Cur)对HepG2细胞组蛋白去乙酰化酶HDAC1的作用及细胞周期依赖激酶抑制剂P21WAF1/CIP1表达的影响,探讨Cur抗肿瘤的作用机制。方法:培养人肝癌细胞株HepG2,用Cur在不同浓度(6.25,12.5,25,50,100μmol.L-1)和不同时间(4,8,16,24,48 h)点处理细胞,提取细胞总蛋白和mRNA,Western blot蛋白质印迹技术检测HDAC1和P21WAF1/CIP1蛋白表达,RT-PCR技术检测P21WAF1/CIP1mRNA的表达水平。结果:Cur对HepG2细胞的半数抑制率(IC50)为25μmol.L-1。IC50浓度作用4 h,HDAC1表达开始降低,降低作用呈时间依赖性;12.5μmol.L-1,24 h可引起HDAC1水平降低,但在50~100μmol.L-1时HDAC1的降低未见明显差别。Cur作用8 h时可见到P21WAF1/CIP1蛋白和mRNA均增加;作用效应呈明显时间和剂量依赖性。结论:Cur明显抑制HDAC1活性和表达,促进P21WAF1/CIP1蛋白和mRNA水平升高。抑制HDAC1活性和促进P21WAF1/CIP1增加可能是Cur抗肿瘤的机制之一。Objective： To investigate the effect of cureumin （Cur） on histone deacetylase （HDAC1） and P21^WAF1/CIP1, a cyclin dependent kinase inhibitor, in HepG2 cells for exploring the mechanism of Cur in anti-cancer. Method： The HDAC1, P21^WAF1/CIP1 proteins and P21^WAF1/CIP1 mRNA were extracted from human hepatoma cells treated with or without Cur of different concentrations at different time points. Western blot analysis was performed to determine the levels of HDAC1 and P21^WAF1/CIP1 proteins, respectively. RT-PCR was performed to detect the level of P21^WAF1/CIP1 mRNA. Result： The IC50 of concentration treated by Cur was 25 μmol·L^-1 on HepG2 cell. The level of HDAC1 was obviously inhibited by Cur, and decreased at 4 hours at IC50 and lasted for 48 h in a time-dependent manner. The inhibition of HDAC1 was significant at the Cur concentration of 12. 5 μmol·L^-1 but there was no difference between 50 and 100 μmol·L^-1. The levels of P21^WAF1/CIP1 mRNA and protein were up-regulated by Cur in dose and time-dependent manner, and the change of mRNA and protein was detected at 8 hours and lasted for 48 hours. Conclusion： Cur can inhibit the level of HDAC1 and enhance the expression of P21^WAF1/CIP1 protein and mRNA, and the results suggest that inhibiting HDAC1 and increasing P21^WAF1/CIP1 may be one of the possible mechanisms of anti-cancer by Cur.

关 键 词：姜黄素 组蛋白去乙酰化酶 细胞周期依赖激酶抑制剂 HEPG2细胞 

分 类 号：R285.5[医药卫生—中药学]