中南大学学报（医学版）
中南大學學報（醫學版）
중남대학학보（의학판）
JOURNAL OF CENTRAL SOUTH UNIVERSITY (MEDICAL SCIENCES)
2009年
5期
369-374
,共6页
乳腺癌%多药耐药%基因%甲基化%组蛋白%乙酰化
乳腺癌%多藥耐藥%基因%甲基化%組蛋白%乙酰化
유선암%다약내약%기인%갑기화%조단백%을선화
breast cancer%multidrug resistance%gene%methylation%histone%acetylation
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DNMT3b mRNA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SIRT1 mRNA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.
目的:分析MCF-7/Adr及MCF-7細胞mdr-1基因啟動子甲基化和組蛋白乙酰化狀態,初步探討乳腺癌多藥耐藥的錶觀遺傳機製. 方法:用甲基化敏感PCR技術檢測兩箇細胞繫mdr-1基因啟動子甲基化狀態.實時定量PCR技術檢測DNA甲基轉移酶(DNA methyltransferases,DNMTs) mRNA及組蛋白去乙酰化酶(histone deacetylases,HDACs) mRNA的錶達.光密度值法檢測組蛋白H3和H4乙酰化水平. 結果:MCF-7細胞mdr-1基因啟動子呈現高甲基化,MCF-7/Adr細胞mdr-1基因啟動子呈現低甲基化.與MCF-7細胞比較,MCF-7/Adr細胞DNMT1,DNMT3a及DNMT3b mRNA錶達顯著下降(P＜0.05).MCF-7/Adr細胞組蛋白H3和H4乙酰化水平較MCF-7細胞明顯升高(P＜0.01).與MCF-7細胞比較,MCF-7/Adr細胞HDAC1,HDAC2,HDAC7及SIRT1 mRNA的錶達顯著下降(P＜0.01). 結論:mdr-1基因啟動子低甲基化、組蛋白H3和H4高乙酰化、DNMTs mRNA及HDACs mRNA低錶達可能是介導MCF-7/Adr細胞MDR形成的重要錶觀遺傳學因素.
목적:분석MCF-7/Adr급MCF-7세포mdr-1기인계동자갑기화화조단백을선화상태,초보탐토유선암다약내약적표관유전궤제. 방법:용갑기화민감PCR기술검측량개세포계mdr-1기인계동자갑기화상태.실시정량PCR기술검측DNA갑기전이매(DNA methyltransferases,DNMTs) mRNA급조단백거을선화매(histone deacetylases,HDACs) mRNA적표체.광밀도치법검측조단백H3화H4을선화수평. 결과:MCF-7세포mdr-1기인계동자정현고갑기화,MCF-7/Adr세포mdr-1기인계동자정현저갑기화.여MCF-7세포비교,MCF-7/Adr세포DNMT1,DNMT3a급DNMT3b mRNA표체현저하강(P＜0.05).MCF-7/Adr세포조단백H3화H4을선화수평교MCF-7세포명현승고(P＜0.01).여MCF-7세포비교,MCF-7/Adr세포HDAC1,HDAC2,HDAC7급SIRT1 mRNA적표체현저하강(P＜0.01). 결론:mdr-1기인계동자저갑기화、조단백H3화H4고을선화、DNMTs mRNA급HDACs mRNA저표체가능시개도MCF-7/Adr세포MDR형성적중요표관유전학인소.
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 methyltransferases (DNMTs) and histone deacetylases (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 mdr-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.
