丙戊酸对体外培养牛胎儿成纤维细胞周期的影响  

Effect of Valproic Acid(VPA) on Cell Growth in Bovine Fibroblast Cells

作　　者：高海霞[1] 王振飞[1] 韩鹏勇[1] 王会敏[1] 王杰[1] 于海泉[1] 

机构地区：[1]内蒙古大学哺乳动物生殖生物学及生物技术教育部重点实验室,呼和浩特010021

出　　处：《农业生物技术学报》2013年第2期223-229,共7页Journal of Agricultural Biotechnology

基　　金：转基因生物新品种培育重大专项(No.2008ZX08007-002)

摘　　要：丙戊酸(valproic acid,VPA)作为一种组蛋白去乙酰化酶抑制剂,能提高细胞内组蛋白乙酰化水平,影响基因的表达。为探讨VPA对体细胞生长的影响,本实验以牛(Bos taurus)胎儿成纤维细胞为研究对象,利用流式细胞技术探讨VPA处理牛体细胞对其细胞周期的影响,并利用Real-timePCR检测VPA处理后细胞转录因子Oct4、Nanog和Cdx2基因表达变化的影响。结果显示,体细胞经0.8、1.0和2.0mmol/L的VPA分别处理24、48和72h后,随VPA浓度的增加,体细胞增殖减缓,细胞周期抑制在间期0/间期1(G0/G1)期。对转录因子Oct4、Nanog和Cdx2基因表达量的检测表明,与未处理组相比VPA处理后细胞Oct4和Nanog的表达量提高,而Cdx2基因的表达量降低。研究结果表明,VPA处理能够改变成纤维细胞的生长特性和基因表达状态,本研究牛体细胞核移植胚胎表观重编程及发育研究提供了参考资料。Valproic acid （VPA） as a histone deacetylase inhibitor, can specifically inhibit the histone deacetylase activity and increase histone acetylation, and can affect gene expression in somatic cells. In this research, the effect of VPA on bovine fetal fibroblast cell growth, cell cycle and gene expression were studied. The bovine （Bos taurus） fetal fibroblasts cells were treated with different concentrations （0.8, 1.0 and 2.0 mmol/L） of VPA for 24, 48 and 72 h, respectively, then cell proliferation and cell cycle were examined by flow cytometry, and the expression of transcriptional factors such as octamer binding factor 4 （Oct4）,Nanog homeobox （Nanog）, and caudal-related homeodomain protein 2 （Cdx2） were detected by Real-time PCR. The results showed that cell growth gradually decelerated with the increase of VPA concentration, cells were mostly arrested at gap0/gap 1 （G0/G 1） phase after treatment. The expression of Oct4 and Nanog genes were improved, while Cdx2 gene expression was reduced. This study showed that VPA treatment affected cell growth, cell cycle, and gene expression of bovine fetal fibroblast cells, which could be benefit to the study on reprogramming of somatic cell after nuclear transfer and embryo development.

关 键 词：VPA 细胞周期 成纤维细胞 转录因子 

分 类 号：Q813[生物学—生物工程]