NS_398调节HepG2细胞组蛋白乙酰化和p21WAF1/CIP1表达

doi:10.3969/j.issn.1004-616x.2007.05.007

癌变·畸变·突变 ›› 2007, Vol. 19 ›› Issue (5): 370-373.doi: 10.3969/j.issn.1004-616x.2007.05.007

NS_398调节HepG2细胞组蛋白乙酰化和p21WAF1/CIP1表达

吴　青/ 朱长才/ 郭晓鹏/ 范丽蓉/ 宋世震

(武汉科技大学医学院，湖北　武汉　430000)

收稿日期:2006-12-11 修回日期:2007-03-12 出版日期:2007-09-30 发布日期:2007-09-30
通讯作者: 宋世震

Regulation of Histone Acetylation and the Expression of p21WAF1/CIP1 by NS_398 in HepG2 Cells

WU Qing,ZHU Chang_cai, GUO Xiao_peng, FAN Li_rong, SONG Shi_zhen

(School of Medicine, Wuhan University of Science and Technology Wuhan 430080,Hubei,China)

Received:2006-12-11 Revised:2007-03-12 Online:2007-09-30 Published:2007-09-30
Contact: SONG Shi_zhen

摘要/Abstract

摘要： 背景与目的：研究非甾体药物NS_398对人肝癌细胞株HepG2细胞组蛋白H3 乙酰化水平的调节作用及对细胞周期素依赖性激酶抑制p21WAF1/CIP1 表达的影响。材料与方法：用不同浓度(100、200、300、400 μmol/L)的NS_398处理 HepG2细胞,以四甲基偶氮唑蓝(MTT)法测定肿瘤细胞增殖抑制率, 流式细胞仪(FCM)检测细胞周期的改变及凋亡百分率的变化,应用NS_398分别作用HepG2细胞4、8、12、24、48 h, 非药物作用组作为对照，提取细胞的总RNA 和总蛋白,采用RT_PCR 技术检测p21WAF1/CIP1 mRNA 表达情况, 并用免疫印迹技术(Western blot) 观察组蛋白H3 的乙酰化水平变化及p21WAF1/CIP1蛋白的表达水平。结果： NS_398抑制HepG2细胞增殖，且呈剂量依赖性,并诱导其凋亡。且呈浓度依赖性改变细胞周期的分布,一方面增高G0/G1期细胞比例，另一方面降低S期和G2/M期细胞比例,与对照组相比差异具有统计学意义(P<0.05)。NS_398对组蛋白H3 乙酰化作用随时间改变而变化,可引起组蛋白H3的乙酰化。NS_398 对p21WAF1/CIP1 mRNA和p21WAF1/CIP1蛋白表达的影响呈时间依赖性。结论： NS_398明显上调HepG2细胞组蛋白H3的乙酰化水平,促进细胞周期依赖性激酶抑制剂p21WAF1/CIP1的表达。

关键词: NS_398, 组蛋白乙酰化, p21WAF1/CIP1, HepG2细胞

Abstract: BACKGROUND ＆ AIM: To investigate the regulation of histone H3 acetylation and the expression of p21WAF1/CIP1 by NS_398 in HepG2 cells. MATERIALS AND METHODS: The effect of NS_398 on the proliferation of HepG2 cells was evaluated by MTT. The apoptotic cells were determined by flow cytometric(FCM)analysis.Total proteins and mRNA were extracted from HepG2 cells treated with or without NS_398 with IC50 200 μmol/L at various time points (4,8,12,24,48 h).By using Western blot, the levels of acetylated histone H3 and p21WAF1/CIP1 protein expression were assayed. RT_PCR was used to detect the expression level of p21WAF1/CIP1 mRNA. RESULTS: NS_398 inhibited cell proliferation and induced apoptosis in HepG2 in a concentration_dependent manner. DNA ploidy analysis showed that S phase cells were significantly decreased with increasing NS_398 concentration.Histone H3 acetylation was obviously increased by NS_398 in a time_dependent manner. The expression levels of p21WAF1/CIP1 mRNA and protein were up_regulated in a time_and dose_dependent manner. CONCLUSION: The level of acetylated histone H3 was up_regulated by NS_398, and the expression of p21WAF1/CIP1 was increased at the same time.

Key words: NS_398, histone acetylation, p21WAF1/ CIP1, HepG2 cell

中图分类号:

Q756

吴　青/ 朱长才/ 郭晓鹏/ 范丽蓉/ 宋世震. NS_398调节HepG2细胞组蛋白乙酰化和p21WAF1/CIP1表达[J]. 癌变·畸变·突变, 2007, 19(5): 370-373.

WU Qing, ZHU Chang_cai, GUO Xiao_peng, FAN Li_rong, SONG Shi_zhen. Regulation of Histone Acetylation and the Expression of p21WAF1/CIP1 by NS_398 in HepG2 Cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2007, 19(5): 370-373.

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NS_398调节HepG2细胞组蛋白乙酰化和p21WAF1/CIP1表达

Regulation of Histone Acetylation and the Expression of p21WAF1/CIP1 by NS_398 in HepG2 Cells

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