干扰Sirt2促进C2C12成肌细胞分化  

Silencing of Sirt2 Promotes C2C12 Myoblast Differentiation

作　　者：吴国芳[1] 路宏朝[1] 贾龙[1] 宋成创 史新娥[1] 杨公社[1] 孙世铎[1] 

机构地区：[1]西北农林科技大学动物脂肪沉积与肌肉发育实验室,陕西杨凌712100

出　　处：《中国生物化学与分子生物学报》2014年第7期714-720,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金(No.31272410);西北农林科技大学"创新团队建设计划"(No.ZD2012007);国家生猪产业体系(No.CARS-36-04B)项目~~

摘　　要：Sirt2是组蛋白去乙酰化酶(HDAC III)家族成员之一,对细胞周期、自噬、脂肪细胞分化、神经细胞存活等生物学过程的调节发挥重要作用.目前,Sirt2在肌肉发育过程中的研究尚未见报道.本文通过构建Sirt2慢病毒干扰载体,侵染C2C12成肌细胞,并用细胞免疫荧光化学、real-time PCR和Western印迹方法,检测其对成肌分化标志基因及相关信号通路因子的影响.结果显示,干扰质粒shRNA-663处理C2C12细胞后,Sirt2 mRNA及蛋白质表达水平与对照相比显著下调(P<0.01);C2C12细胞分化第4 d,MyoD,MyoG,MyHC mRNA及蛋白质表达均显著增加(P<0.01);PI3K,AKT,FoxO1磷酸化水平明显升高.结果表明,Sirt2可通过PI3K/AKT/FOXO1信号通路来促进成肌细胞分化,是肌生成的一个潜在调节因子.Sirt2 is one of the members of histone deacetyltransferase （HDAC III） family, which plays a keyrole in the regulation of various cellular processes, including cell cycle, autophagy, neuroprotection, adiposedifferentiation. Now, the study of Sirt2 on muscle development has not been reported. The effect of Sirt2 onmuscle differentiation was investigated by constructing Sirt2 lentivirus vectors to treat C2C12 myoblast, and wealso used immunofluorescent staining, real-time PCR and Western blotting to evaluate related gene expressionlevels. The results showed that the expression levels of Sirt2 mRNA and protein in C2C12 myoblast treated bySift2 lentivirus plasmid were significantly lower than those of control group. At day 4 of C2C12 myoblastdifferentiation, the expression of MyoD, MyoG and MyHC mRNA and protein as well as the phosphorylation ofPI3K, AKT and FoxO1 were significantly increased. It was suggested that Sirt2 could promote myoblastdifferentiation via the PI3K/AKT/FOXO1 signal pathway and be a potential regulator of myogenesis.

关 键 词：Sirt2 成肌分化 C2C12 干扰载体 

分 类 号：Q782[生物学—分子生物学]