大鼠肝再生的肝细胞干性变化中性别决定区转录因子2 mRNA和其他非编码RNA的表达变化与作用

Expression change and role of sex determining region box transcription factor 2 mRNA and the non-coding RNA in the hepatocyte stem changes during the rat liver regeneration

目的了解大鼠肝再生(LR)0 h和2 h时性别决定区转录因子2(SOX2)基因及其mRNA相互作用的微小RNA(miRNA,miR)和环状RNA(circRNA)的表达变化、相互作用和调节肝细胞干性变化的途径与方式。方法按Higgins等方法制备大鼠2/3肝切除(partial hepatectomy,PH)模型,按Smedsrod等方法分离肝细胞,用大规模定量检测技术测定大鼠肝再生中肝细胞的mRNA、miRNA合称内源竞争RNA(ceRNA)的表达变化,用Cytoscape 3.2软件构建ceRNA的相互作用网,用ceRNA综合分析等方法解析它们的表达相关性和作用相关性。结果PH后0 h和2 h时,SOX2 mRNA的比值为1.00±0.09和2.15±0.48,miR-3558-3p为4.53±0.10和0.81±0.16,circRNA_18404为1.24±0.04和11.10±0.57,circRNA_18045为1.97±0.47和4.44±0.23。同时,PH后0 h时,SOX2促进的富含AT的交互域5A(ARID5A)、激活转录因子3(ATF3)、BTG抗增殖因子2(BTG2)等8个细胞去分化相关基因表达下调,抑制的细胞分化相关基因干扰素调节因子6(IRF6)和生长抑素(SST)表达上调。PH后2 h时,SOX2促进的ARID5A、ATF3、BTG2等8个细胞去分化相关基因表达上调,抑制的细胞分化相关基因SST表达下调、IRF6未发生有意义表达变化。结论上述circRNA抑制的miRNA、miRNA抑制的SOX2 mRNA以及SOX2调节的细胞干性相关基因的表达相关性和作用相关性有利于PH后0 h时肝细胞处于分化状态和PH后2 h时肝细胞处于干性状态。

Objective To explore the role pathway and pattern of the sex determining region box transcription factor 2(SOX2)and its mRNA interaction with microRNA(miRNAs,miR)and circular RNA(circRNA)at 0 hour and 2 hours in the rat liver regeneration.Methods The rat 2/3 hepatectomy(partial hepatectomy,PH)model was prepared as described by Higgins,the hepatocytes were isolated according to the method of Smedsrod et al,the expression changes of mRNA,miRNA and circRNA[together named as competing endogenous RNAs(ceRNA)]were detected by the large-scale quantitative detection technology,the interaction network of ceRNA was constructed by Cytoscape 3.2 software,and their correlation in expression and role were analyzed by ceRNA comprehensive analysis.Results It was found that at the 0 hour and 2 hours after PH,the ratio value of SOX2 mRNA shows 1.00±0.09 and 2.15±0.48,miR-3558-3p displays 4.53±0.10 and 0.81±0.16,circRNA_18404 shows 1.24±0.04 and 11.10±0.57,circRNA_18045 displays 1.97±0.47 and 4.44±0.23.At the same time,the eight kinds of cell dedifferentiation-related genes AT-rich interaction domain 5A(ARID5A),activating transcription factor 3(ATF3),BTG anti-proliferation factor 2(BTG2),etc,which are prometed in expression by SOX2,were down-regulated at 0 h after PH,but the cell differentiation-related genes interferon regulatory factor 6(IRF6)and somatostatin(SST),which are inhibited in expression by SOX2,were up-regulated at 0 hour after PH.On the other hand,the eight kinds of cell dedifferentiation-related genes ARID5A,ATF3,BTG2,etc,which are promoted in expression by SOX2,were up-regulated at 2 hours after PH,but the cell differentiation-related gene SST,which is inhibited in expression by SOX2,was down-regulated,and IRF6 had no meaningful changes in expression at 2 hours after PH.Conclusion The correlation in expression and role of the miRNA,which are inhibited by circRNA,SOX2,its mRNA is inhibited by miRNA,and the cell stem-related genes,which are regulated by SOX2,are helpful for the hepatocyte to be in differentiation state at 0 hour after PH and to be in stem state at 2 hours after PH.