组蛋白H3K9甲基修饰酶在2周、4周和6周龄小鼠肝脏中的表达  

Expressions of histone H3K9 methylation modified enzymes in the liver of two-week,four-week and six-week old mice

作　　者：郭晓强[1,2,3] 陆菁潇[2] 王越甲[1] 张巧霞[2] 蔡志明[2] 常彦忠[1] 段相林[1] 

机构地区：[1]河北师范大学生命科学学院,河北省动物生理生化与分子生物学重点实验室,石家庄050016 [2]北京大学深圳医院男性生殖与遗传广东省重点实验室,广东深圳518036 [3]解放军白求恩军医学院生化教研室,石家庄050081

出　　处：《解剖学报》2011年第5期661-665,共5页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(38070265);河北省自然科学基金资助项目(C2010000410)

摘　　要：目的探讨不同周龄小鼠肝脏组蛋白H3K9甲基转移酶(KMT)和去甲基化酶(KDM)的表达。方法取2周、4周和6周龄BALB/c雄性小鼠肝脏组织,分别提取总mRNA和蛋白质,应用实时定量反转录-聚合酶链式反应(RT-qPCR)确定几种H3K9甲基转移酶和去甲基化酶的表达差异,使用Western blotting检测两种H3K9去甲基化酶KDM3A和KDM4B的蛋白含量。结果几种H3K9甲基转移酶(包括KMT1C、KMT1E和PRDM2)和去甲基化酶(包括KDM3A、KDM3B、KDM4A和KDM4B)的mRNA在小鼠不同年龄阶段的表达存在差异(P<0.05,每年龄段5只小鼠),而两种组蛋白去甲基化酶KDM3A和KDM4B在蛋白水平也有明显不同,6周龄小鼠肝脏中两种蛋白表达明显低于2周和4周(P<0.05,每年龄段5只小鼠)。结论特定基因位点的H3K9甲基化状态与肝脏发育和生物学功能可能存在密切联系。Objective To investigate the expressions of H3K9 methyhransferases （KMTs） and demethylases （KDMs） in the liver of different aged mice. Methods Two-week, four-week and six-week old mice were killed and their liver tissues collected. Both mRNA and protein from the mice liver were extracted using Trizol kit and sonicated method respectively. Reverse transcription-quantitative real time polymerase chain reaction （RT-qPCR） was performed for mRNA expressions of several kinds of H3K9 methyhransferases and demethylases. The levels of H3K9 demethylases KDM3A and KDM4B proteins were determined using Western blotting. Results The mRNA expressions of several KMTs（ including KMT1C, KMT1E and PRDM2 ） and KDMs（including KDM3A, KDM3B, KDM4A and KDM4B） were obviously different in the different developmental stages of a mouse＇ s liver（P 〈 0. 05, n = 5）. The levels of KDM3A and KDM4B proteins in the mice liver were also distinct, and both proteins in six-week old mice were lower than those in two-week and four-week old mice （P 〈 0. 05, n = 3）. Conclusion The H3K9 methylation status of specific gene loci may be closely associated with liver development and biological function.

关 键 词：组蛋白 H3K9甲基转移酶 H3K9去甲基化酶 肝脏 免疫印迹法 实时定量聚合酶链式反应 小鼠 

分 类 号：R730.231[医药卫生—肿瘤]