TGF-β2对心肌细胞H9C2转录因子表达的影响和其组蛋白H3乙.doc

TGF-β2对心肌细胞H9C2转录因子表达的影响及其组蛋白H3乙酰化调控机制 王文文1,2，田 杰1，郑 敏1,2，孙慧超1，陈国珍2 [400014重庆，重庆医科大学附属儿童医院：心血管内科1，儿童发育疾病研究省部共建教育部重点实验室，儿科学重庆市重点实验室，重庆市（儿童发育重大疾病诊治与预防）国际科技合作基地2] [摘要] 目的 探讨转化生长因子β2（transforming growth factor beta2TGF-β2）心肌细胞H9C2Mef2c和GATA4组蛋白乙酰化的。方法 TGF-β2对细胞的影响。利用Real-time PCR检测心肌细胞的心脏相关转录因子Mef2c和GATA4 mRNA的相对表达量，筛选合适的TGF-β2干预浓度。采用Western blot 检测心肌细胞乙酰化H3（cetylated histone H3，acH3）蛋白的表达差异。（hromatin immunoprecipitation，CIP）Real-time PCRMef2c和GATA4acH3结合DNA的相对表达水平。比色分析法检测TGFβ2对心肌细胞的总体组蛋白乙酰化酶（HATs）活性的影响。结果 ①TGFβ2干预组相对于对照组心肌细胞的增殖能力增强（P0.05）。②心肌细胞的心脏相关转录因子Mef2c和GATA4 mRNA的相对表达量随着TGF-β2浓度的升高呈现先升高后降低的趋势，在5μg/的浓度时均达到最高，ef2c mRNA相对表达量是对照组的1.8倍，GATA4 mRNA是对照组的2.3倍（P0.0）。TGF-β2干预72h后acH3蛋白表达量是对照组的4倍（P 0.05），HATs活性是对照组的1.34倍（P 0.05）。TGF-β2干预组Mef2c和GATA42.56和2.16倍（P 0.05）。结论 5μg/ TGF-β2可以促进心肌细胞H9C2的分裂与增殖，心脏相关转录因子Mef2c和GATA4的表达Mef2c和GATA4表达[关键词] TGFβ2；心肌细胞H9C2；组蛋白H3；乙酰化；表观遗传学 [中图法分类号] R725.4 [文献标志码]Transforming growth factor beta2 up-regulates transcription factors in myocardial cell H9C2 by acetylation of histone H3 Wang Wenwen1,2 , Tian Jie1, Zheng Min1,2, Sun Huichao1, Chen Guozhen2 (1Department of Cardiology, 2Key Laboratory of Developmental Diseases in Childhood of Ministry of Education, Chongqing International Science and Technology Cooperation Center for Child Development and Disorders, Chongqing Key Laboratory of Pediatrics, Children’s Hospital, Chongqing Medical University, Chongqing, 400014, China) [Abstract] Objective To determine the effect of transforming growth factor beta2 (TGF-β2) on histone acetylation of myocardial cell strain H9C2 and its role in regulating the expression of cardiac development-related transcription factors. Methods H9C2 cells were cultured and then treated with TGF-β2 at concentrations of 1.25 , 2.5, 5, 10 or 20 μg/L. Cell morphology was observed by inverted phase contrast microscopy (IPCM), and cell counting was carried out at 12, 24, 48 and 72 h after treatment to draw the growth curve. Quantitative real-