5-氮脱氧胞苷及曲古抑菌素A影响供核细胞H19基因表达的时间依赖性及浓度依赖性  

Time- and concentration-dependent effect of 5-aza-2'-deoxycytidine and trichostain A on H19 gene expression in nuclear donor cells

作　　者：杨晓煜[1,2,3] 姚建凤[1,2] 黄燕芳[4] 张更[1] 徐榕清[1] 郑鸣[1] 

机构地区：[1]福建医科大学基础医学院人体解剖学与组织胚胎学系,福州350004 [2]福建医科大学细胞与发育工程研究中心,福州350004 [3]福建省生殖医学(工程)中心,福州350004 [4]福建医科大学附属第一医院体检中心,福州350005

出　　处：《第二军医大学学报》2008年第12期1451-1454,共4页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(30500282);福建省高等学校新世纪优秀人才支持计划(NCETFJ-0603);福建省青年人才创新项目(2005J044)~~

摘　　要：目的:探讨甲基转移酶抑制剂——5-氮脱氧胞苷(5-aza-2′-deoxycytidine,5-aza-dC)和组蛋白去乙酰基酶抑制剂——曲古抑菌素A(trichostatin A,TSA)影响供核细胞H19基因表达的时间依赖性和浓度依赖性。方法:用不同浓度5-aza-dC及TSA按不同作用时间处理供核细胞(鼠尾尖成纤维样细胞),利用实时定量PCR法检测处理后H19基因的表达。结果:不同浓度的5-aza-dC处理组之间H19的表达无明显差异,但延长作用时间可使H19表达增强;TSA使H19的表达下降,但无时间依赖性。结论:长时间低浓度的5-aza-dC能增强供核细胞中H19基因的表达,而TSA则使H19的表达减弱,为体细胞克隆中表观遗传修饰抑制剂的选择及后续基因印迹研究提供了依据。Objective：To study the time- and concentration-dependent effect of DNA methyhransferase inhibitor 5-aza-2＇- deoxycytidine（5-aza dC） and histone deacetylase inhibitor triehostatin A（TSA） on the expression of H19 gene in nuclear donor cells. Methods： Donor cells （fibroblast-like cells from tail tip of mice） were treated with 5-aza-dC and TSA at different concentrations for different time periods. The expression of H19 was determined by real-time PCR after treatment. Results： The expression of H19 was not significantly different between groups treated with different concentrations of 5-aza-dC, but the expression of H19 was increased with the increase of treatment time. TSA decreased H19 expression, hut not in a time-dependent manner. Conclusion： Long time exposure to low concentration of 5-aza-dC can increase the expression of H19; TSA can decrease the expression of H19 ,which provides a basis for selection of epigenetic modification inhibitor and later study on imprinting in somatic cell nuclear transfer.

关 键 词：H19基因 甲基转移酶抑制剂 组蛋白脱乙酰基酶抑制剂 5-AZA-DC TSA 供核细胞 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学]