MSCs心肌样细胞分化过程中GCN5募集促分化相关蛋白的筛选

发布时间：2018-10-05 10:59

【摘要】：筛选并分析间充质干细胞(mesenchymal stem cells,MSCs)诱导分化为心肌样细胞过程中GCN5招募促分化相关蛋白集合体的组成。免疫荧光细胞化学、实时荧光定量PCR鉴定MSCs经5-氮杂胞苷(5-azacytidine,5-azaC)诱导分化为心肌样细胞;免疫共沉淀技术分离、串联质谱鉴定GCN5募集蛋白集合体组成,并从心肌细胞分化角度筛选、验证分化相关蛋白因子。MSCs经5-azaC诱导分化的心肌样细胞表达心肌特异性基因GATA4、MEF2C和心肌细胞结构、功能蛋白cTnt、MHC和Cx43;筛选、验证出心肌细胞分化相关GCN5招募蛋白归类为:(1)DNA结合蛋白Sp1/KLF;(2)转录辅激活子PGC-1α和Rb1;(3)转录延伸复合体组成成分以及信号通路蛋白Akt。通过筛选获得MSCs经诱导向心肌样细胞分化过程中心肌分化相关蛋白因子,为进一步研究干细胞分化信号传导途径、特异性生物靶点干预以及提高干细胞分化效率打下了基础。
[Abstract]:To screen and analyze the composition of GCN5 recruitment associated protein aggregates for differentiation into cardiomyocyte-like cells induced by mesenchymal stem cells (mesenchymal stem cells,MSCs). Immunofluorescence cytochemistry and real-time fluorescence quantitative PCR were used to identify the differentiation of MSCs into cardiomyocyte-like cells induced by 5-azacytidine (5-azacytidine 5-azaC), co-immunoprecipitation technique, tandem mass spectrometry (MS) to identify the composition of GCN5 recruitment protein aggregates, and screening from the perspective of cardiomyocyte differentiation. To verify the expression of cardiomyocyte specific gene GATA4,MEF2C and cardiac myocyte structure by 5-azaC induced by differentiation related protein factor. The functional protein cTnt,MHC and Cx43; were screened. GCN5 recruitment proteins associated with cardiomyocyte differentiation were classified as: (1) DNA binding protein Sp1/KLF; (2) transcriptional coactivators PGC-1 伪 and Rb1; (3) transcription extension complex components and signal pathway protein Akt. Through screening, we obtained the related protein factor of myocardial differentiation in the process of MSCs induced cardiomyocyte differentiation, which laid a foundation for further study on the signal transduction pathway of stem cell differentiation, the intervention of specific biological target and the improvement of stem cell differentiation efficiency.
【作者单位】：重庆医科大学附属儿童医院儿科研究所;重庆医科大学附属儿童医院心血管内科;
【基金】：国家自然科学基金资助项目(No.30973219)~~
【分类号】：R329

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