BMSCs向心肌分化的基因表达谱分析  

DYNAMIC CHANGES OF GENE EXPRESSION PROFILES DURING CARDIOMYOGENESIS OF HUMAN MARROW MESENCHYMAL STEM CELLS

作　　者：杨琳[1] 沈悌[2] 陈连凤[2] 曹欣欣[2] 赖晋智[3] 

机构地区：[1]华中科技大学同济医学院附属同济医院肿瘤科,武汉430030 [2]中国医学科学院北京协和医学院北京协和医院血液内科 [3]中国医学科学院北京协和医学院北京协和医院心内科

出　　处：《中国修复重建外科杂志》2012年第5期607-611,共5页Chinese Journal of Reparative and Reconstructive Surgery

摘　　要：目的分析hBMSCs经5-氮杂胞苷(5-azacytidine,5-aza)诱导向心肌样细胞分化过程中基因表达谱的改变。方法取胸外科非血液病患者手术中废弃的肋骨骨髓分离培养hBMSCs,5-aza诱导第2代hBMSCs。取诱导前及诱导后的细胞,免疫细胞化学法检测α-actin、肌钙蛋白T(cardiac troponin T,cTnT)和连接蛋白43(connexin 43)表达,流式细胞仪检测cTnT阳性细胞百分率;利用人表达谱基因芯片技术筛选分化过程中的差异表达基因,对部分基因进行功能分类和分层聚类分析。结果 hBMSCs经5-aza诱导后,部分呈肌细胞样形态。免疫细胞化学染色检测示诱导前细胞α-actin、cTnT染色呈弱阳性,connexin 43染色呈阴性;诱导后3周细胞α-actin、cTnT、connexin 43染色均呈阳性。流式细胞仪检测示诱导前cTnT阳性细胞百分率为7.43%±0.02%,诱导后3周为49.64%±0.05%。分化过程中共检测到1 814个显著差异表达基因,对其中647个基因分层聚类,聚为5类,生物功能包括信号传导、细胞代谢、增殖分化、发育以及形态发生等。结论 hBMSCs经5-aza诱导向心肌样细胞分化过程受信号传导通路、转录基因、生长因子等多种因素在不同时间点上的共同调控。Objective To analyze the changes of gene expression profiles during the process that human bone marrow mesenchymal stem cells （hBMSCs） are induced to differentiate into cardiomyogenic cells with 5-azacytidine （5- aza）. Methods hBMSCs were isolated from marrow of obsolete ribs and induced with 5-aza. Then immunocytochemical staining was used to detect the expressions of ct-actin, cardiac troponin T （cTnT）, and connexin 43, and the percentage of cTnT positive cells was tested with flow cytometry. In the process of differentiation, variation of gene expression was screenedwith Genechips Operating System of human gene expression profiles. And the differentially expressed genes were functionally analyzed and hierarchical clustered. Results When BMSCs were induced in vitro with 5-aza, part of the cells turned into myogenic cells morphologically. Before induction, immunocytochemical staining for α-actin and cTnT showed slight positive and for connexin 43 showed negative. While after 3 weeks of induction, immunocytochemical staining for α-actin, cTnT, and connexin 43 showed all positive. With flow cytometry, the percentage of cTnT positive cells was 7.43% ± 0.02% before induction, but it was 49.64% ± 0.05% after induction. During differentiation, 1 814 differentially expressed genes were reported by gene chips. Of them, 647 genes were divided into 5 groups with hierarchical clustering. They had various biological functions, involving signal transduction, cell metabolism, proliferation, differentiation, development, and topogenesis. Conclusion hBMSCs can differentiate into cardiomyogenic cells with the induction of 5-aza in vitro. Multiple genes related with signal transduction, transcription, and growth factors are involved during this process.

关 键 词：HBMSCS 基因表达谱 诱导 心肌样细胞 

分 类 号：R346[医药卫生—基础医学]