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姜黄素对人淋巴瘤细胞系Raji中组蛋白H3乙酰化的影响

[Effect of curcumin on acetylation of histone H3 in human lymphoma cell line Raji].

作者信息

Li Xin-Gang, Chen Yan, Wu Qing, Sun Chun-Yan

机构信息

Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430022, P. R. China.

出版信息

Ai Zheng. 2006 May;25(5):582-6.

Abstract

BACKGROUND & OBJECTIVE: Epigenetic change is an important mechanism of oncogenesis. The inhibitors of methyltransferases and deacetylases, with epigenetic modificative effects, could inhibit proliferation and induce apoptosis of tumor cells. This study was to investigate the effects of curcumin on the acetylation of histone H3 and the expression of p21(WAF1/CIP1) gene in human lymphoma cell line Raji.

METHODS

Raji cells were treated with 25 micromol/L curcumin. The levels of acetylated histone H3 and p21WAF1/CIP1 were detected by Western blot, the expression of p21(WAF1/CIP1) gene was detected by reverse transcription-polymerase chain reaction (RT-PCR), and the level of acetylated histone H3 at the site of p21(WAF1/CIP1) promoter gene was examined by chromatin immunoprecipitation assay. Cell cycle distribution was studied by flow cytometry.

RESULTS

Curcumin induced hyperacetylation of histone H3 at the site of p21(WAF1/CIP1) promoter by 1.9 folds, and enhanced the levels of p21(WAF1/CIP1) mRNA by 4.2 folds and protein by 5.1 folds 24 h after treatment. Raji cells were arrested at G(2)/M phase when treated with curcumin for 24 h, and at G(0)/G(1) phase when treated for 36 h.

CONCLUSION

Curcumin, with epigenetic modificative effects, could enhance the acetylayion of histone H3 at the site of p21(WAF1/CIP1) promoter gene, improve transcription of p21(WAF1/CIP1) gene, and arrest cell cycle progression of Raji cells.

摘要

背景与目的

表观遗传改变是肿瘤发生的重要机制。具有表观遗传修饰作用的甲基转移酶和去乙酰化酶抑制剂可抑制肿瘤细胞增殖并诱导其凋亡。本研究旨在探讨姜黄素对人淋巴瘤细胞系Raji中组蛋白H3乙酰化及p21(WAF1/CIP1)基因表达的影响。

方法

用25μmol/L姜黄素处理Raji细胞。采用蛋白质免疫印迹法检测乙酰化组蛋白H3和p21WAF1/CIP1的水平,逆转录-聚合酶链反应(RT-PCR)检测p21(WAF1/CIP1)基因的表达,染色质免疫沉淀法检测p21(WAF1/CIP1)启动子基因位点的乙酰化组蛋白H3水平。通过流式细胞术研究细胞周期分布。

结果

姜黄素处理24小时后,使p21(WAF1/CIP1)启动子基因位点的组蛋白H3乙酰化水平提高了1.9倍,p21(WAF1/CIP1)mRNA水平提高了4.2倍,蛋白质水平提高了5.1倍。用姜黄素处理Raji细胞24小时,细胞停滞于G(2)/M期;处理36小时,细胞停滞于G(0)/G(1)期。

结论

具有表观遗传修饰作用的姜黄素可增强p21(WAF1/CIP1)启动子基因位点的组蛋白H3乙酰化,促进p21(WAF1/CIP1)基因转录,阻滞Raji细胞的细胞周期进程。

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