【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。更多还原

【Abstract】 Objective To investigate the expressions of H3K9 methyltransferases(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 methyltransferases 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.更多还原