[1]王文文,田杰,郑敏,等.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[J].第三军医大学学报,2013,35(07):639-643.
 Wang Wenwen,Tian Jie,Zheng Min,et al.Transforming growth factor beta2 up-regulates transcription factors in myocardial cell H9C2 by acetylation of histone H3[J].J Third Mil Med Univ,2013,35(07):639-643.
点击复制

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(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
35卷
期数:
2013年第07期
页码:
639-643
栏目:
论著
出版日期:
2013-04-15

文章信息/Info

Title:
Transforming growth factor beta2 up-regulates transcription factors in myocardial cell H9C2 by acetylation of histone H3
作者:
王文文田杰郑敏孙慧超陈国珍
重庆医科大学附属儿童医院:心血管内科,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
Author(s):
Wang Wenwen Tian Jie Zheng Min Sun Huichao Chen Guozhen
Department of Cardiology, Key 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
关键词:
TGF-β2心肌细胞H9C2组蛋白H3乙酰化表观遗传学
Keywords:
transforming growth factor beta2 myocardial cell H9C2histone H3acetylation epigenetics
分类号:
R322.11; R329.28; R394.3
文献标志码:
A
摘要:
目的      探讨转化生长因子-β2(transforming growth factor beta2,TGF-β2)对心肌细胞H9C2转录因子Mef2c和GATA4的影响及其与组蛋白H3乙酰化的关系。      方法       比较TGF-β2对细胞生长的影响。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组蛋白H3(acetylated histone H3,acH3)的表达差异。染色质免疫共沉淀(chromatin immunoprecipitation,ChIP)Real-time PCR检测Mef2c和GATA4启动子区与acH3结合DNA的相对表达水平。比色分析法检测TGF-β2对心肌细胞的总体组蛋白乙酰化酶(HATs)活性的影响。      结果       ①TGF-β2干预组相对于对照组心肌细胞的增殖能力增强(P<0.05)。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5 μg/L的浓度时均达到最高,Mef2c mRNA相对表达量是对照组的1.8倍,GATA4 mRNA是对照组的2.3倍(P<0.05)。③TGF-β2干预72 h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)。④TGF-β2干预组Mef2c和GATA4启动子区组蛋白H3的乙酰化水平分别是对照组的2.56和2.16倍(P<0.05)。      结论       5 μg/L TGF-β2可以促进心肌细胞H9C2的分裂与增殖,并上调心脏相关转录因子Mef2c和GATA4的表达, 组蛋白H3乙酰化水平的升高可能是Mef2c和GATA4表达上调的机制之一。
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-time PCR assay was used to select suitable intervention concentration of TGF-β2 by comparing the mRNA levels of Mef2c and GATA4. Expression of acetylated histone H3 (acH3) was assessed by Western blot assays. Histone Acetylase (HATs) Activity Colorimetric Assay Kit was used to measure the level of HATs activities. And, chromatin immunoprecipitation (ChIP) real-time PCR was employed to measure the level of acH3 in promoter regions of Mef2c and GATA4.        Results        TGF-β2  enhanced the proliferation in H9C2 cells(P<0.05). After TGF-β2 treatment, the mRNA expression of Mef2c and GATA4 was elevated in a dose-dependent manner, and then reached peak at a dose of 5 μg/L with an increase by 1.8 folds and 2.3 folds, respectively(P<0.05). The acetylation level of histone H3 increased by 4-fold (P<0.05), meanwhile, the HATs activity increased by 1.34-fold (P<0.05). TGF-β2 treatment resulted in the expression level of acH3 enhanced by 2.56- and 2.16-fold respectively in the promoter regions of Mef2c and GATA4 (P<0.05).        Conclusion       TGF-β2 at a dose of 5 μg/L promotes the proliferation in H9C2 cells, and up-regulates the expression of Mef2c and GATA4, which might be through improving the acetylation of histone H3.

参考文献/References:

王文文, 田杰, 郑敏, 等. 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[J].第三军医大学学报,2013,35(7):639-643.

更新日期/Last Update: 2013-04-07