Islet-1在乙酰化调控网络中特异性辅助C3H10T1/2细胞向心肌样细胞分化  

Islet-1 specifically assists the process of C3H10T1/2 cell differentiating into cardiomyocyte-like cells in the histone acetylation regulatory network

作　　者：林建萍[1] 田杰[1,2] 刘官信[1] 鲁荣[1] 刘建平[1] 智深深[1] 朱静[1] 

机构地区：[1]重庆医科大学附属儿童医院儿科研究所儿童发育与疾病研究教育部重点实验室,重庆400014 [2]重庆医科大学附属儿童医院心脏内科,重庆400014

出　　处：《基础医学与临床》2012年第4期363-368,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(No.30973219);重庆市自然科学基金(No.2009BA5048)

摘　　要：目的筛选并分析转染Islet-1慢病毒载体的C3H10T1/2细胞转化为心肌样细胞过程中与Islet-1相互作用的组蛋白乙酰化酶(HATs)和组蛋白去乙酰化酶(HDACs),明确Islet-1在C3H10T1/2细胞分化为心肌样细胞乙酰化调控网络中的关键枢纽作用。方法培养转染Islet-1慢病毒载体的C3H10T1/2细胞,观察细胞形态。免疫荧光和免疫印迹检测Islet-1的表达部位和最高表达时间点。免疫共沉淀与Islet-1结合的蛋白。免疫印迹验证Islet-1相互作用的HATs和HDACs。结果诱导组细胞形态出现心肌样细胞改变。各组Islet-1主要在胞质表达。诱导组Islet-1表达量在诱导后3周最高(0.782±0.015)。诱导组Islet-1表达量显著高于空白对照组和C3H10组(分别为0.819±0.026,0.127±0.006和0.126±0.001)(P<0.05),免疫共沉淀技术可行。与Islet-1相互作用的HATs和HDACs有GCN5、P300/CBP和HDAC4。结论 Islet-1与GCN5、P300/CBP和HDAC4相互作用特异性辅助C3H10T1/2细胞向心肌样细胞分化。Objective To find out which protein domains of histone acetylase（HATs） and histone deacetylase（HDACs） interact with Islet-1 during the differentiation of C3H10T1/2 cells transfected with Islet-1 lentiviral vector into cardiomyocyte-like cells,and to prove the pivotal role of Islet-1 in the network of histone acetylation.MethodsCulture and observe C3H10T1/2 cells transfected with Islet-1 lentiviral vector.The position of Islet-1 was detected by Immunofluorescence cytochemistry.The protein extraction time was determined by Western-blot according to expression levels of Islet-1-immunoprecipitation.The Islet-1-binding protein was precipitated by co-immunoprecipitation（Co-IP）.The proteins of HATs and HDACs interacting with Islet-1 were verified by Western-blot.ResultsCells in induction group showed the appearance of cardiomyocyte-like cells.The location of Islet-1 was located in cytoplasm.The expression of protein in induction group was significant in the third week（0.782±0.015） compared with the first 2 weeks.The expression of Islet-1 in induction group was significantly higher than that in blank control group and C3H10 group（0.819±0.026,0.127±0.006 and 0.126±0.001 respectively,P0.05）.The proteins interacting with Islet-1 were GCN5、P300/CBP and HDAC4.Conclusions The synergism of Islet-1 and its interacting proteins such as GCN5、P300/CBP and HDAC4 played an important role in the differentiation of C3H10T1/2 cells into cardiomyocyte-like cells.

关 键 词：C3H10T1/2细胞 Islet-1 心肌样细胞 组蛋白乙酰化 

分 类 号：R363[医药卫生—病理学]