组蛋白去乙酰化酶抑制剂对人胚胎成纤维细胞表观遗传的影响  

Effect of histone deacetylase inhibitor on epigenetic change of human embryonic fibroblasts

作　　者：徐小明[1,2] 段馨[1] 周欢群[1] 林戈[1,2] 卢光琇[1,2] 

机构地区：[1]中南大学生殖与干细胞工程研究所,湖南长沙410008 [2]中南大学人类干细胞国家工程研究中心,湖南长沙410008

出　　处：《西北农林科技大学学报（自然科学版）》2011年第8期29-33,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家自然科学基金项目(30800650);湖南省自然科学基金项目(09JJ4010)

摘　　要：【目的】探讨组蛋白去乙酰化酶抑制剂（曲古菌素A（TSA）和丙戊酸盐（VPA））对人胚胎成纤维细胞（human embryonic fibroblasts,hEF）组蛋白乙酰化（acH4K12）和组蛋白单甲基化（H4K20me1）的影响,为进一步探讨组蛋白去乙酰化酶抑制剂对人类体细胞核移植胚胎表观遗传重编程的作用机制奠定基础。【方法】使用0400 nmol/L TSA或04 mmol/L VPA处理hEF 24 h,用核型分析及荧光原位杂交（FISH）检测hEF细胞遗传学变化,使用间接免疫荧光染色和激光共聚焦显微镜观察TSA和VPA对hEF的acH4K12及H4K20me1水平的影响。【结果】TSA处理组hEF细胞呈现出形态学变化,且TSA具有细胞毒性作用。VPA处理组与未处理组细胞无明显的形态学变化和细胞毒性作用。核型分析及FISH检测结果显示,TSA和VPA处理组与未处理组均维持正常的核型（46,XX）,未发生明显的细胞遗传学改变。间接免疫荧光染色结果表明,acH4K12和H4K20me1水平随着TSA和VPA浓度的增加而升高。【结论】TSA和VPA可以提高hEF的acH4K12和H4K20me1水平,且未发生明显的细胞遗传学变化。【Objective】 The study was conducted to explore the effect of histone deacetylase inhibitor（TSA and VPA） on epigenetic change of human embryonic fibroblasts（hEF）,and to lay the foundation for further investigating the histone deacetylase inhibitor on the epigenetic mechanisms for reprogramming of human somatic cell nuclear transfer（SCNT） embryos.【Method】 The hEF were treated with 0-400 nmol/L TSA or 0-4 mmol/L VPA for 24 h.Cytogenetic change was analysed with karyotype analysis and fluorescence in situ hybridization（FISH）.After hEF were treated with TSA or VPA,histone acetylation（acH4K12） and methylation（H4K20me1） were examined by using indirect immunofluorescence staining and scanning confocal microscopy.【Result】 There was difference between group of TSA treatment and group of non-TSA treatment in cell morphous change and TSA had cytotoxic effect.However,there was no obvious difference between group of VPA treatment and group of non-VPA treatment in cell morphous change and VPA had no cytotoxic effect.Groups of TSA or VPA treatment and groups of non-TSA or non-VPA treatment all kept normal karyotype（46,XX） and without obvious cytogenetic abnormality with karyotype analysis and FISH dectection.The results of indirect immunofluorescence staining showed that the levels of acH4K12 and H4K20me1 were increased with increasing concentrations of TSA and VPA treatment.【Conclusion】 TSA and VPA can increase the levels of acH4K12 and H4K20me1 in hEF and have no obvious cytogenetic changes.

关 键 词：组蛋白去乙酰化酶抑制剂 表观遗传 成纤维细胞 

分 类 号：Q813.7[生物学—生物工程]