转卵泡抑制素基因对绵羊肌源性细胞中 肌肉抑制素基因的调控

doi:10.3864/j.issn.0578-1752.2013.02.018

Abstract

Abstract: 【Objective】In this research, skeletal muscle specific expressed follistatin（FST）eukaryotic expression vectors, pCFCDs-red and pSPFCDs-red, were constructed, then respectively transfected into different cell lines. FST and downstream gene expression situation in RNA and protein levels was detected, and the simultaneous start efficiency of skeletal muscle-specific promoter of sub-SP and α-actin was assayed, so that to understand the effect of specific expression of FST on the development of muscle.【Method】Using liposomes, the eukaryotic expression vectors of pCFCDs-red and pSPFCDs-red were transfected respectively into sheep fetal fibroblast cells (SFFCs), skeletal muscle satellite cells (SMSCs) and SMSCs induced muscle tube, and the transgenic cells were analyzed on status of cell growth, cell cycle, cell size and expression of target gene and downstream gene respectively by flow cytometry, Real-Time quantitative PCR and western blotting. 【Result】The growth of transgenic cells basically comply with the trend of non-transgenic cells, and transgenic SMSCs cell had a higher proliferation rates than that genetically modified SFFC. Flow cytometry analysis showed that more than 70% of cells of transgenic cell lines were in G0/G1 phase, maintained a strong ability to divide and had a single peak ,a good aneuploidy and volume increased significantly. Real-time quantitative PCR and western blotting results showed that RNA and protein levels of FST in the transgenic cell lines were raised compared to non-transgenic cell lines. Transgenic SMSCs and myotubes compared to transgenic SFFCs cell had higher expression of FST. Transfected pSPFCDs-red transgenic cells had higher expression level of FST than transfected pCFCDs-red cells.【Conclusion】Skeletal muscle specific promoter SP and α-actin can effectively start FST target gene in the SMSCs and myotubes, and FST have a higher expression level in myotubes. Start efficiency of SP promoter is higher than that of α-actin. In myogenic cells, the increased FST can cause the reduction of MSTN proteins level. FST has a certain role in promoting growth and development of skeletal muscle cells. Key words: FST; synthetic promoter SP ;specific promoter α-actin; skeletal muscle satellite cell; myotubes

Key words: FST , synthetic promoter SP , specific promoter α-actin , skeletal muscle satellite cell , myotubes

YUE Qun-Hua, YUAN Jian-Long, HAO Fei, JIN Mu-Zi, WANG Jing-Yuan, YANG Wen-Liang, LIU Dong-Jun, CANG Ming. Transfected FST Gene into Ovine Muscle-Drived Myogenic Cells to Regulate the Expression of MSTN Gene[J].Scientia Agricultura Sinica, 2013, 46(2): 385-393.

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Transfected FST Gene into Ovine Muscle-Drived Myogenic Cells to Regulate the Expression of MSTN Gene

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