【摘要】 探讨低表达丝切蛋白CFL2基因对小鼠成肌细胞C2C12信号通路关键成分的影响。采用RNAi技术,将猪CFL2基因真核表达shRNA重组体稳定转染至小鼠成肌细胞C2C12后提取其总RNA和总蛋白质,采用Real-time PCR和Western blot检测CFL2基因低表达后成肌细胞C2C12信号通路关键成分(MEF2C、sk MLCK、Rho A、AMPKα2、p38和Ca M)的表达情况。结果表明:在3个稳转细胞株中MEF2C、Ca M、p38和AMPKα2的mRNA表达均为极显著下调(P<0.01),sk MLCK的mRNA表达为极显著上调(P<0.01),其它成分不显著;MEF2C和Ca M的蛋白表达在3个稳转细胞株中均为显著或极显著下调(P<0.05或P<0.01),其他4种成分(p38、Rho A、AMPKα2和sk MLCK)均差异不显著(P>0.05),说明CFL2低表达显著下调信号通路关键成分MEF2C和Ca M的表达,CFL2与Ca M和sk MLCK分别呈显著正相关和负相关。更多还原

【Abstract】 To investigate the effect of cofilin CFL2 low expression on the key factors in signal pathway of muscle fiber,mouse myoblast C2C12 was transfected by shRNA recombinant vectors targeting CFL2 gene according to RNAi method.The mRNA and protein expression levels of CFL2 and the key factors of signal pathway(MEF2 C,sk MLCK,Rho A,AMPKα2,p38 and Ca M) in the low expression groups were detected by Real-time PCR and Western blot.Results showed that in the low expression groups,the mRNA levels of key factors in signal pathway such as MEF2 C,Ca M,P38 and AMPKα2 were down-regulated and sk MLCK was increased significantly(P<0.01).The protein levels for key factors MEF2 C and Ca M in three transfected cell strains were down-regulated significantly(P<0.05 or P<0.01).The other four factors(p38,Rho A,AMPKα2 and sk MLCK) showed no significant difference(P > 0.05).It suggests that CFL2 gene silencing caused the low expression of MEF2 C and Ca M.There was a significantly positive or negative correlation between CFL2 and Ca M or sk MLCK.更多还原