MCF-7/Adr细胞mdr-1基因启动子甲基化和组蛋白乙酰化状态与多药耐药的关系(英文)  

Relation of promoter methylation of mdr-1 gene and histone acetylation status with multidrug resistance in MCF-7/Adr cells

作　　者：黄程辉[1] 曹培国[1] 谢兆霞[2] 

机构地区：[1]中南大学湘雅三医院肿瘤科,长沙410013 [2]中南大学湘雅医院血液科,长沙410078

出　　处：《中南大学学报（医学版）》2009年第5期369-374,共6页Journal of Central South University ：Medical Science

基　　金：supported by Natural Science Foundation of Hunan Province,P.R.China (07JJ4009)

摘　　要：目的:分析MCF-7/Adr及MCF-7细胞mdr-1基因启动子甲基化和组蛋白乙酰化状态,初步探讨乳腺癌多药耐药的表观遗传机制。方法:用甲基化敏感PCR技术检测两个细胞系mdr-1基因启动子甲基化状态。实时定量PCR技术检测DNA甲基转移酶(DNA methyltransferases,DNMTs)mRNA及组蛋白去乙酰化酶(histone deacetylases,HDACs)mRNA的表达。光密度值法检测组蛋白H3和H4乙酰化水平。结果:MCF-7细胞mdr-1基因启动子呈现高甲基化,MCF-7/Adr细胞mdr-1基因启动子呈现低甲基化。与MCF-7细胞比较,MCF-7/Adr细胞DNMT1,DNMT3a及DNMT3bmRNA表达显著下降(P<0.05)。MCF-7/Adr细胞组蛋白H3和H4乙酰化水平较MCF-7细胞明显升高(P<0.01)。与MCF-7细胞比较,MCF-7/Adr细胞HDAC1,HDAC2,HDAC7及SIRT1mRNA的表达显著下降(P<0.01)。结论:mdr-1基因启动子低甲基化、组蛋白H3和H4高乙酰化、DNMTs mRNA及HDACs mRNA低表达可能是介导MCF-7/Adr细胞MDR形成的重要表观遗传学因素。Objective To analyze the mdr-1 gene promoter methylation and histone acetylation status in both MCF-7/Adr and MCF-7 cells and to preliminarily explore the epigenetic mechanism of multidrug resistance in breast cancer. Methods mdr-1 gene promoter methylation status of the 2 cell lines was detected by methylation-sensitive PCR. mRNA expression of DNA methyhransferases （DNMTs） and histone deaeetylases （HDACs） was detected by real-time quantitative PCR. Acetylation level of histone H3 and H4 was examined by optical density assay. Results Promoter hypermethylation of mdr-1 gene was observed in MCF-7 cells whereas hypomethylation was found in MCF-7/Adr cells. Expression of DNMT1 ,DNMT3a, and DNMT3b mRNA in MCF-7/Adr cells significantly decreased compared with that of MCF-7 cells （ P 〈 0. 05 ）. H3 and H4 histone acetylation levels of MCF-7/Adr cells were obviously higher than those of the MCF-7 cells （P 〈 0.01 ）. Expression of HDAC1, HDAC2, HDAC7, and Sirtuin type 1 （SIRT1） mRNA in MCF-7/Adr cells was significantly reduced （ P 〈 0.05 ）. Conclusion Hypomethylation of the promoter region of rndr-1 gene, high H3 and H4 histone acetylation, and low mRNA expression of DNMTs and HDACs may be important epigenetic factors for the development of MDR in MCF- 7/Adr cells.

关 键 词：乳腺癌 多药耐药 基因 甲基化 组蛋白 乙酰化 

分 类 号：R737.9[医药卫生—肿瘤] R965[医药卫生—临床医学]