Hes1基因诱导小鼠肝原始细胞分化为胆管上皮细胞

背景与目的： 采用体外获得性表达外源发状分裂相关增强子-1(hairy and enhancer of split 1,Hes1)基因的方法，探讨Hes1在肝干细胞分化以及胆管上皮细胞发育中的作用。 材料与方法： 通过PCR方法从小鼠基因组中克隆Hes1基因片段，构建表达载体pEGFP-C1-Hes1和pcDNA3.1- Hes1，将2种表达载体分别转染肝原始细胞系(LEPCs) ，应用RT-PCR和Real-time PCR技术检测胆管细胞分子标志物CK19、GGT，胆管上皮细胞相关转录因子HNF6、HNF1β，肝细胞分子标志物GS、BGP和肝卵圆细胞的分子标志物Thy-1的表达，并在荧光显微镜下观察EGFP标记的LEPCs细胞系荧光强度的变化。结果：成功构建表达载体pEGFP-C1- Hes1和pcDNA3.1-Hes1。RT-PCR和Real-time PCR检测均表明胆管细胞分子标志CK19、GGT表达量上调；胆管上皮细胞相关转录因子HNF6、HNF1β表达上调；肝细胞分子标志GS、BGP表达量下调；卵圆细胞的分子标志Thy-1表达量下调；LEPCs细胞系绿色荧光增强。 结论： 初步证明小鼠肝原始细胞经Hes1的表达诱导后可向胆管上皮细胞方向分化，推测Hes1为胆管上皮细胞分化的转录调控因子。

Hes 1 Induced Differentiation of Mouse Liver Progenitor Cells to Cholangiocytes

BACKGROUND AND AIM:To study the differentiation of mouse liver epithelial progenitor cells(LEPCs)to cholangiocytes induced by hairy and enhancer of split 1(Hes1)gene in vitro.MATERIALS AND METHODS:The Hes1 gene was amplified by PCR from mouse tail genomic DNA and subcloned into pEGFP-C1 and pcDNA3.1 vector.LEPCs were tranfected and performed in vitro.Cholangiocyte markers,CK19,GGT,HNF6 and HNF1B;hepatocyte markers,GS and BGP;hepatic oval cell marker,Thy-1 were assayed with RT-PCR and Real-time PCR.The fluorescence of the LEPCs tagged with EGFP was observed in vitro.RESULTS:Cholangiocyte markers,CK19,GGT,HNF6 and HNF1B, were up-regulated.Hepatocyte markers GS and BGP,and hepatic oval cell marker Thy-1,were down-regulated. CONCLUSION:It indicated that LEPCs differentiated to cholangiocytes initially in vitro.and Hesl might play important mles in the differentiation of LEPCs and influence cholangiocyte versus hepatocyte fate determination.