姜黄素抑制Raji细胞组蛋白去乙酰化酶活性阻止细胞周期进程

Study on the mechanism of arrest cell cycle progression of Raji cells by curcumin

目的:研究姜黄素对Raji细胞组蛋白去乙酰化酶(HDAC1)的抑制作用及细胞周期蛋白p21WAF1/CIP1的表达情况,探讨姜黄素抗淋巴瘤的机制。方法:培养人B细胞淋巴瘤细胞株Raji,应用姜黄素在不同浓度和时间点处理细胞,提取细胞的总蛋白和RNA,用Western blot印迹技术检测HDAC1和p21WAF1/CIP1蛋白的表达,并用RT-PCR技术检测p21WAF1/CIP1mRNA的水平,用流式细胞仪分析细胞周期。结果:(1)姜黄素明显抑制HDAC1的表达,在4h开始下降,持续48h,呈时间和浓度依赖性;(2)姜黄素明显促进p21WAF1/CIP1蛋白和p21WAF1/CIP1mRNA表达,在8h mRNA开始上升,12h可以检测到p21WAF1/CIP1蛋白表达升高,也呈时间和浓度依赖效应;(3)姜黄素在24h时,明显诱导Raji细胞G0/G1期阻止。结论:姜黄素能够抑制HDAC1的表达,并对细胞周期蛋白p21WAF1/CIP1和p21WAF1/CIP1mRNA的表达有明显促进作用,阻止Raji细胞在G0/G1期。抑制HDAC1活性和促进p21WAF1/CIP1表达可能是姜黄素抗淋巴瘤的机制之一。

OBJECTIVE To investigate the effect of curcumin on histone deacetylase-1 （HDAC1）and cell cycle protein p21^WAF1/CIP1 in Raji cells for exploring the basic mechanism of curcumin on anti-lymphoma. METHODS After B-lymphoma cell lines Raji cell cultured and treated with or without curcumin at different concentration（50,25,12. 5,6. 25,3. 125μmol· L^-1 ） for various times（0,4,8, 12,24,48 h）, the total protein and RNA were extracted,Western blot analysis were performed to determine the level of HDAC1 and p21^WAF1/CIP1 respectively,and RT-PCR was performed to dectect the level of p21^WAF1/CIP1 mRNA, and DNA content was determined using flow cytometer to analyse cell cycle. RESULTS （1）The level of HDAC1 was inhibited by curcumin at a dose and time dependent manner,and HDAC1 reduced beginning from 4 hours and lasted 48 hours;（2） The level of p21^WAF1/CIP1 protein and p21^WAF1/CIP1 mRNA were up-regulated at a dose and time dependent manner, and p21^WAF1/CIP1 mRNA were detected as early as 8 hours, p21^WAF1/CIP1 protein increased at 12 hours and lasted 48 hours; （3） Curcumin significantly induced G0/G1 phase arrest following 24 h of exposure 12. 5μmol·L^-1 concentration. CONCLUSION Curcumin is able to inhibit the expression of HDAC1 and promote the expression level of p21^WAF1/CIP1 protein and mRNA,and arrest Raji cells in G0/G1 phase. The results suggest that inhibiting HDAC1 and increasing p21^WAF1/CIP1 may be one possible mechanism of curcumin on anti-lymphoma.