| 摘要: |
| 目的 研究乙酰化酶抑制剂Garcinol对雌激素促乳腺癌细胞MCF-7增殖、细胞周期转化及凋亡抑制的影响及机制。 方法 应用CCK-8法检测Garcinol 对17β-雌二醇（17β-E2）促MCF-7细胞增殖的影响；流式细胞术检测细胞周期和细胞凋亡情况；细胞免疫荧光检测NF-κB/p65核转运情况；RT-PCR检测cyclin D1、Bcl-2、Bcl-xL mRNA水平；蛋白质印迹分析检测乙酰化（ac）-H3、ac-H4、NF-κB/ac-p65、cyclin D1、Bcl-2、Bcl-xL 蛋白的表达水平。 结果 Garcinol能抑制17β-E2促细胞增殖作用，使细胞周期阻滞于G0/G1期，细胞凋亡率增加（P<0.01）；17β-E2促进MCF-7细胞ac-H3、ac-H4、NF-κB/ac-p65表达水平（P<0.01，P<0.05，P<0.01），Garcinol能抑制17β-E2对ac-H3、NF-κB/ac-p65的表达促进作用（P<0.01），对ac-H4影响无统计学意义；17β-E2促进NF-κB/p65核转运相应受到Garcinol抑制（P<0.01）；17β-E2促进cyclin D1、Bcl-2、Bcl-xL的 mRNA转录和蛋白表达水平的作用受到Garcinol抑制（P<0.05）。 结论 17β-E2促MCF-7细胞增殖和凋亡抑制作用与组蛋白和非组蛋白NF-κB/p65乙酰化水平增高有关，乙酰化酶抑制剂Garcinol 通过降低乙酰化水平对雌激素促乳腺癌细胞增殖具有明显抑制作用，降低NF-κB通路的ac-p65蛋白水平，从而下调cyclin D1、Bcl-2、Bcl-xL可能是其重要机制。 |
| 关键词: Garcinol 乙酰化作用 组蛋白类 NF-κB 乳腺肿瘤 |
| DOI：10.3724/SP.J.1008.2014.00714 |
| 投稿时间：2013-12-16修订日期：2014-01-24 |
| 基金项目:国家自然科学基金（30900651） |
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| Effect of acetyltransferase inhibitor Garcinol on estrogen-promoted proliferation of breast cancer cell line MCF-7 and the related mechanism |
| YE Xia,DENG Hua-yu*,ZHANG Li,ZHAO Jing,YUAN Lei |
(Department of Pathophysiology, Laboratory for Stem Cell and Tissue Engineering, College of Basic Medicine, Chongqing Medical University, Chongqing 400016, China
*Corresponding author.) |
| Abstract: |
| Objective To investigate the effect of acetyltransferase inhibitor Garcinol on estrogen-induced proliferation, cell cycle promotion and apoptosis inhibition of human breast cancer MCF-7 cells and the related mechanism. Methods The effect of Garcinol on 17β-estradiol (17β-E2)-induced proliferation of MCF-7 cells was investigated using CCK-8 assay, and the optimal concentration of Garcinol was determine according to the inhibition rate. The cell cycle and apoptosis were analyzed by flow cytometry; the nuclear translocation of NF-κB/p65 was examined by immune cell fluorescence. RT-PCR was used to determine the expressions of cyclin D1, Bcl-2 and Bcl-xL mRNA in MCF-7 cells; and the expressions of ac-H3, ac-H4, NF-κB/ac-p65, cyclin D1, Bcl-2, and Bcl-xL protein were determined by Western blotting analysis. Results Acetyltransferase inhibitor Garcinol inhibited 17β-E2-induced proliferation of MCF-7 cells, arrested MCF-7 cell cycle at G0/G1 phase, and increased the cell apoptosis(P<0.01). 17β-E2 increased the expressions of ac-H3, ac-H4 and NF-κB/ac-p65 protein in MCF-7 cells(P<0.01, P<0.05, P<0.01), while Garcinol significantly inhibited the increase of ac-H3 and NF-κB/ac-p65 (P<0.01) but not ac-H4(P>0.05). 17β-E2-induced nuclear translocation of NF-κB/p65 in MCF-7 cells was also significantly inhibited by Garcinol (P<0.01). Garcinol also significantly inhibited 17β-E2-induced transcription and protein expression of cyclin D1, Bcl-2 and Bcl-xL mRNA in MCF-7 cells (P<0.05). Conclusion 17β-E2-induced proliferation and apoptosis inhibition of MCF-7 cells are associated with elevated acetylation level of histone and nonhistone NF-κB/p65, and acetyltransferase inhibitor Garcinol may inhibit the effect of 17β-E2 by decreasing acetylation, probably through decreasing ac-p65 expression of NF-κB pathway, and subsequently down-regulating cyclin D1,Bcl-2,and Bcl-xL. |
| Key words: Garcinol acetylation histones NF-κB breast neoplasms |
