Expression of Klf4 and Nanog and their promoter methylation during differentiation of rat bone marrow CD90+Lin-cells to hepatocytes
LIU Qincheng
ZHANG Zilang
PAN Runhua
LIU Ning
HAN Xiaoyu
GUO Linghong
WANG Chunming
LIAO Caixian
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.
Keywords：bone marrow mesenchymal stem cellsbone marrow CD90+Lin-cellsNanog geneKlf4 gene
Publication Date：2017-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 13-16 )