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作 者:黎冠宏[1] 王薇[1] 李继业[1] 阿斯楞[2] 潘润华[1] 廖彩仙[1]
机构地区:[1]南方医科大学南方医院肝胆外科,广东广州510515 [2]内蒙古医科大学附属医院普外科,内蒙古呼和浩特010050
出 处:《山东大学学报(医学版)》2014年第6期28-32,共5页Journal of Shandong University:Health Sciences
基 金:广东省科技计划(2010B080701069)
摘 要:目的研究大鼠骨髓源性肝干细胞(BMDLSC)向肝细胞诱导分化过程中H3K27二甲基化变化情况,初步探讨H3K27二甲基化与BMDLSC向肝细胞分化过程的关系。方法分离收集大鼠骨髓细胞,通过免疫磁珠分选获得BMDLSC,而后体外诱导其分化为肝细胞,RT-PCR动态检测其白蛋白(ALB)及甲胎蛋白(AFP)的mRNA表达水平,通过Western blotting及免疫荧光染色检测第0、7、14天核小体组蛋白H3K27二甲基化的水平。结果分离纯化的BMDLSC在体外诱导分化过程中ALB的表达水平逐渐升高,而AFP的表达水平则先升高后下降;新鲜分离的BMDLSC H3K27无明显甲基化,诱导分化后第7天H3K27二甲基化水平较前升高,第14天进一步升高。结论 BMDLSC经体外诱导后定向分化为肝细胞,在分化过程中H3K27二甲基化水平逐渐升高,H3K27二甲基化修饰变化可能参与BMDLSC细胞向肝细胞分化的调控。Objective To explore the changes of H3K27 dimethylation during the differentiation of bone marrow-derived liver stem cells ( BMDLSC) into hepatocytes from rats in vitro and to investigate the relationship between them. Methods BMDLSCs collected were isolated from rat bone marrow using a magnetic activated cell sorting ( MACS) . Purified BMDLSCs were induced into hepatocytes in vitro.The expressions of albumin ( ALB) and alpha fetoprotein ( AFP) were detected dynamically by RT-PCR.The changes of histone H3K27 dimethylation were measured by West-ern blotting and immunofluorescence staining on day 0, 7, and 14.Results The expression of ALB gradually increased when BMDLSCs were induced in vitro, while AFP expression firstly rose and later fell.H3K27 from fresh BMDLSCs showed no significant methylation at first, but on day 7 after differentiation, the level of H3K27 dimethyla-tion significantly increased and further rose on day 14.Conclusion The level of dimethylation shows a gradual increase during the differentiation of BMDLSCs.H3K27 dimethylation may regulate the differentiation of BMDLSC into hepatocytes.
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