山东医药2017，Vol.57Issue(31)：13-16,4.DOI:10.3969/j.issn.1002-266X.2017.31.004
大鼠CD90+Lin-骨髓细胞诱导分化为肝细胞过程中Klf4、Nanog基因的表达及启动子区甲基化观察
Expression of Klf4 and Nanog and their promoter methylation during differentiation of rat bone marrow CD90+Lin-cells to hepatocytes
摘要
Abstract
Objective To observe the expression of Klf4 and Nanog and the promoter region CpG site methylation changes during differentiation of rat bone marrow CD90+Lin-cells to hepatocytes.Methods Rat bone marrow CD90+Lin-cells were induced in vitro by 25 μg/L recombinant human hepatocyte growth factor and 0.1 nmol/L dexamethasone.DNA and RNA were extracted from cells on day 0, 7, and 14 of induction.Real-time fluorescent quantitative PCR was used to detect Klf4 and Nanog mRNA expression.Methylation Specific PCR was performed to detect methylation variation of CpG sites in Klf4 and Nanog promoters.Results Compared with the day 0, the relative expression of Klf4 mRNA decreased on day 7 and day 14 (P<0.05);compared with the day 0, the relative expression of Nanog mRNA decreased on day 7 and day 14 (P<0.05).Compared with the day 0, the methylation frequency of the first and second CpG sites in Klf4 promoter increased on day 7 (P<0.05);compared with the day 7, the methylation frequency of the first CpG site in Klf4 promoter increased on day 14, while the second and third CpG sites in Klf4 promoter decreased (P<0.05).Compared with the day 0, the methylation frequency of the first site in Nanog promoter decreased in day 7, while the second sites in Nanog promoter increased (P<0.05);compared with the day 7, the methylation frequency of the first CpG site in Nanog promoter increased on day 14, while the second CpG site in Nanog promoter decreased (P<0.05).Conclusions During differentiation of rat bone marrow CD90+Lin-cells to hepatocytes in vitro, the expression of Klf4 and Nanog first increases and then decreases as time passes, and the CpG sites methylated degrees of both promoters vary as time passes.The methylation of CpG site in the promoter region of Klf4 and Nanog not only affects the expression of Klf4 and Nanog, but also plays an important regulatory role.关键词
骨髓间充质干细胞/CD90+Lin-骨髓细胞/Nanog基因/Klf4基因Key words
bone marrow mesenchymal stem cells/bone marrow CD90+Lin-cells/Nanog gene/Klf4 gene分类
医药卫生引用本文复制引用
刘钦成,张梓朗,潘润华,刘宁,韩晓玉,郭凌宏,王春明,廖彩仙..大鼠CD90+Lin-骨髓细胞诱导分化为肝细胞过程中Klf4、Nanog基因的表达及启动子区甲基化观察[J].山东医药,2017,57(31):13-16,4.基金项目
广东省科技计划项目(2010B080701069). （2010B080701069）
