Effect of MSTN Propeptide and shRNA Co-expression Vector on Proliferation of Skeletal Muscle Satellite Cells  

作　　者：Feng Lin-he Wang Xin Lu Ming Tong Hui-li Li Shu-feng Yan Yun-qin 

机构地区：[1]College of Life Sciences, Northeast Agricultural University

出　　处：《Journal of Northeast Agricultural University(English Edition)》2014年第1期31-38,共8页东北农业大学学报（英文版）

基　　金：Supported by the Major Special Projects of New Product Training of Transgenic Organisms(zx080072008-2008)

摘　　要：Myostatin （MSTN） is a negative regulator of skeletal muscle growth, in order to study the effect of inhibition MSTN expression on the proliferation of bovine skeletal muscle satellite cells, we constructed co-expression vector pcDNA3.1-Pro- MSTNshRNA, transfected it into muscle satellite cells by Liposome 2000, and detected cell proliferation changes by CCK-8 method and flow cytometry after 48 h. The expressions of P21 and CDK2 were detected by Western blot and real-time PCR. The results showed that the cell vitality of experimental groups significantly increased than that of the negative control, and cells in S phase also increased significantly （P〈0.05）. After knocked down MSTN gene, P21 expression decreased （P〈0.05）, but CDK2 gene expression increased （P〈0.05）. These results indicated that MSTN gene expression was associated with P21 and CDK2, the proliferation of skeletal muscle satellite cells could be promoted while MSTN was inhibited, which provided a theoretical basis for the study on transgenic cattle.Myostatin （MSTN） is a negative regulator of skeletal muscle growth, in order to study the effect of inhibition MSTN expression on the proliferation of bovine skeletal muscle satellite cells, we constructed co-expression vector pcDNA3.1-Pro- MSTNshRNA, transfected it into muscle satellite cells by Liposome 2000, and detected cell proliferation changes by CCK-8 method and flow cytometry after 48 h. The expressions of P21 and CDK2 were detected by Western blot and real-time PCR. The results showed that the cell vitality of experimental groups significantly increased than that of the negative control, and cells in S phase also increased significantly （P〈0.05）. After knocked down MSTN gene, P21 expression decreased （P〈0.05）, but CDK2 gene expression increased （P〈0.05）. These results indicated that MSTN gene expression was associated with P21 and CDK2, the proliferation of skeletal muscle satellite cells could be promoted while MSTN was inhibited, which provided a theoretical basis for the study on transgenic cattle.

关 键 词：MYOSTATIN cell proliferation flow cytometry expression vector 

分 类 号：Q291[生物学—细胞生物学]