Guo Linghong, Jiang Huiyuan, Lu Yanjun, Liu Maoxi, Liu Haiyi
Colorectal Surgery, Shanxi Cancer Hospital Taiyuan 030013, Shanxi, China.
Am J Transl Res. 2023 Dec 15;15(12):6718-6726. eCollection 2023.
To investigate the changes and functions of Sox2 gene expression and promoter methylation during induced differentiation of bone marrow mesenchymal stem cells (BMSCs) into hepatocytes (HCs).
Rat bone marrow Thy-1Lin cells were prepared and divided into control group (directed induction of differentiation into HCs) and experimental group (5-azacytidine intervention induced differentiation). The mRNA expression levels of ALB and Sox2 were detected by fluorescence quantitative polymerase chain reaction (PCR), and the Sox2 gene promoter methylation level was determined by Bisulfite sequencing PCR (BSP).
Sox mRNA expression level was significantly increased in experimental group compared to the control group at 0, 7, and 14 days, respectively (all P<0.05). The Sox2 promoter methylation level was gradually increased after 0, 7 and 14 days induction in both groups, accompanied by an increase in methylated loci (all P<0.05). Statistical significance was present in CpG methylated loci between groups (all P<0.05).
The expression of Sox2 gene increased first and then decreased in the process of inducing rat BMSCs into stem cells, and the methylation level of CpG loci in the promoter region changed dynamically, with an increased overall methylation level. After 5-aza treatment, the Sox2 promoter was in a non-methylated state, and its mRNA expression increased, which hindered the cell differentiation.
探讨骨髓间充质干细胞（BMSCs）向肝细胞（HCs）诱导分化过程中Sox2基因表达及启动子甲基化的变化和作用。
制备大鼠骨髓Thy-1Lin细胞，分为对照组（定向诱导分化为HCs）和实验组（5-氮杂胞苷干预诱导分化）。采用荧光定量聚合酶链反应（PCR）检测ALB和Sox2的mRNA表达水平，采用亚硫酸氢盐测序PCR（BSP）检测Sox2基因启动子甲基化水平。
实验组在0、7和14天时Sox mRNA表达水平分别较对照组显著升高（均P<0.05）。两组在诱导0、7和14天后Sox2启动子甲基化水平逐渐升高，同时甲基化位点增加（均P<0.05）。组间CpG甲基化位点存在统计学差异（均P<0.05）。
在大鼠BMSCs诱导分化为干细胞的过程中，Sox2基因表达先升高后降低，启动子区域CpG位点甲基化水平动态变化，总体甲基化水平升高。5-氮杂胞苷处理后，Sox2启动子处于非甲基化状态，其mRNA表达增加，阻碍了细胞分化。
