肌肉生长抑素基因过表达对3T3-L1细胞增殖的影响

Influence of myostatin over-expression on the proliferation of 3T3-L1 preadipocytes

目的观察小鼠肌肉生长抑素(mMSTN)对小鼠3T3-L1前脂肪细胞增殖的影响。方法小鼠肌肉组织mMSTN基因表达序列,构建pcDNA3.1(+)-mMSTN真核表达质粒。采用阳离子脂质体转染法,观察mMSTN过表达对细胞增殖的影响。结果 DNA测序鉴定证实成功构建mMSTN真核表达质粒。质粒转染后,mMSTN mRNA和蛋白表达显著增高,分别达到对照组的7.63倍和1.38倍(P值均<0.05),而且能明显促进3T3-L1前脂肪细胞的增殖,是对照组的1.67倍(P<0.001)。结论成功构建mMSTN真核表达载体,该载体过表达显著促进3T3-L1细胞的增殖。

Objective To investigate the influence of mouse myostatin （mMSTN） on the proliferation of 333 - L1 preadipo- cytes. Methods Mouse MSTN full cDNA sequence was amplified by RT - PCR to construct pcDNA3.1 （ ＋ ） - mMSTN ex- pression plasmid. Cationic liposome - mediated transfection was applied to observe the influence of mMSTN over - expression on proliferation of 3T3 - L1 preadipocy-tes. Results DNA sequencing confirmed the successful construction of mMSTN expression plasmid pcDNA3.1 （ - ） - mMSTN. After transfecting mMSTN expression plasmid into 3T3 - L1 preadipocytes, both mMSTN mRNA and protein levels increased significantly and reached 7.63 times and 1.38 times of those in control group（P 〈 0.05 ）, respectively. In addition, MSTN over- expression can significantly stimulate 33＂3 -L1 cells proliferation. The influence of MSTN on proliferation was 1.67 times higher than what was observed in control cells in 72 h following transfection（ P 〈 0. 001 ）. Conclusion mMSTN expression plasmid pcDNA3.1 （ - ） - mMSTN was established and successfully expressed in 333 - L1 preadipocytes both in mRNA and protein levels. Additionally, MSTN can significantly promote 3T3 - L1 cells proliferation.