组蛋白去乙酰化下调对肾细胞癌BNIP3基因表达的影响

目的探讨肾细胞癌中线粒体促凋亡蛋白BNIP3表达下调机制。方法运用CCK-8法及流式细胞技术检测组蛋白去乙酰化酶抑制剂曲古霉素A（TSA）对肾癌细胞株786-O、ACHN、A498增殖、凋亡的影响；运用实时荧光定量PCR（Q-PCR）和蛋白质免疫印记（Western blot）技术检测TSA对肾癌细胞BNIP3表达的影响；运用染色质免疫沉淀（ChIP）技术检测TSA对肾癌细胞BNIP3基因启动子区组蛋白H3乙酰化状态的影响。结果经TSA处理后，3种肾癌细胞增殖均受到显著抑制（P<0.05），细胞早期凋亡明显增加，BNIP3 mRNA（P<0.05）和蛋白表达水平上调。786-O、ACHN的BNIP3基因启动子区组蛋白H3呈去乙酰化状态，TSA恢复了其乙酰化状态；A498的BNIP3基因启动子区组蛋白H3呈乙酰化状态。结论BNIP3基因启动子区组蛋白去乙酰化是其在肾癌中表达下调的主要机制，并可能参与了肾癌的发生发展。

英文摘要:

Objective To investigate the down-regulation mechanism of（bcl-2/adenovirus E1B 19 kDa interacting protein 3 (BNIP3) expression in renal cell carcinoma (RCC). Methods RCC cell lines 786-O，ACHN and A498 were treated with different concentrations of histone deacetylase inhibitor TSA. Thereafter, the proliferation of RCC cells was determined with CCK-8 assay，cell apoptosis was observed by flow cytometry， and the expression levels of BNIP3 were determined by Q-PCR and Western blot，and the acetylation status of histone H3 in the promoter of BNIP3 was detected by ChIP. Results After the treatment with TSA，the proliferation of the three RCC cell lines was significantly inhibited (P<0.05)，the early apoptosis of cells obviously increased， and the expression levels of BNIP3 mRNA (P<0.05) and protein were up-regulated. The histone H3 in BNIP3 promoter of both 786-O and ACHN was deacetylated，while the histone H3 in BNIP3 promoter of A498 was acetylated. Conclusion Histone deacetylation may be the important mechanism of BNIP3 silencing in RCC.

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