牛肌肉抑制素前肽对牛肌卫星细胞增殖的影响

通过PCR扩增目的基因,构建真核表达载体,利用PEI转染牛肌卫星细胞,转染后48与72 h通过CCK-8与流式细胞术检测细胞增殖的变化,通过RT-PCR检测目的基因的表达,通过RT-PCR与Western blot检测下游基因P21,CDK2的表达变化.结果表明,在两个时间点,实验组细胞活力明显升高(P<0.01),G0/G1期细胞所占的比例明显下降(P<0.01),S期细胞所占比例明显上升(P<0.01),MSTN前肽基因有大量表达,P21表达下降,CDK2表达上升.结果揭示,在牛肌卫星细胞中,MSTN前肽通过下调P21,上调CDK2的表达量促进细胞增殖.旨在揭示牛MSTN前肽基因对牛肌卫星细胞(MDSC)增殖的影响,为后续的转基因牛试验提供理论依据.

Effect of bovine myostatin propeptide on bovine muscle satellite cell proliferation

The research intended to reveal the effects of bovine MSTN propeptide gene on bovine muscle satellite cell proliferation,and provided a theoretical basis for subsequent trial of genetically modified cattle.The trial got the objective gene fragment by means of PCR amplification and constructed eukaryotic expression vector.The vector was delivered into muscle satellite cells by PEI.At 48 h and 72 h after transfection,detected the change of cell proliferation by CCK-8 and flow cytometry and detected the expression of the objective gene.The trial detected the expression change of downstream gene P21,CDK2 by RT-PCR and Western blot.The results showed that: at two time points,the vigor of experimental group cells was significantly increased;the proportion of G0/G1 phase cells was significantly decreased;the proportion of S phase cells was significantly increased;the MSTN propeptide had a large number of gene expression;the expression of P21 was reduced;the expression of CDK2 was increased.These resu;ts indicated that MSTN propeptide promoted cell proliferation by down-reguLating the expression of P21 and up-regulating the expression of CDK2 in MDSC.