转胰岛素样生长因子Ⅰ基因促细胞增殖作用的研究  

The Influence of Trans-IGF-1 Gene on Cell Proliferation

作　　者：梁东春[1] 张欣[2] 孙书军[2] 张镜宇[1] 

机构地区：[1]天津医科大学内分泌研究所,300070 [2]天津大学

出　　处：《天津医药》2004年第11期677-678,i001,共3页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(项目编号:50233020)

摘　　要：目的:研究转胰岛素样生长因子1(IGF-1)基因对细胞增殖的影响。方法:构建含IGF-1信号肽及成熟肽全部编码序列的重组报告基因载体pTracer-IGF-1,脂质体法将该重组质粒转染入体外培养的C2C12小鼠成肌细胞。以杀稻瘟毒素(blasticidin)筛选出稳定转染的细胞并进行克隆化,3H-TdR法检测细胞增殖率。结果:与对照组相比,转IGF-1基因细胞的增殖速率明显增加,其培养基亦可显著促进细胞的增殖。结论:所转染的IGF-1基因可在受体细胞内稳定的表达,合成的IGF-1可被分泌至培养基中,在局部发挥促细胞增殖作用。Objective: Test the influence of trans-IGF-1 gene on cell proliferation in vitro. Methods: Firstly a DNA fragment coding both signal and mature peptide of IGF-1 was subcloned into reporter gene plasmid pTracer-CMV/Bsd. Therefore recombinant plasmid pTracer-cIGF-1 was delivered into C2C12 mouse myoblast celline by lipofectamine 2000. Screened steadily transfected cells by selected media containing blasticidin 5 μg/mL, and 3H-TdR method was applied to detect proliferation rate of the cells. Results: 48 h after transfection green areas similar with the shape of cell could be seen under fluroscent microscope. As a result, trans-IGF-1 cell had an more increased proliferation rate than untransfected C2C12 cell and the media of trans-IGF-1 cell could also accelerated the proliferation rate of cultured C2C12 cell markedly. Conclusion: Transfected IGF-1 gene could by expressed steadily in host cell. IGF-1 synthesized by the trans-IGF-1 cell could be secreted out of the cell and could enhance cell proliferation locally.

关 键 词：IGF-1 细胞增殖 报告基因 胰岛素样生长因子Ⅰ 转染 克隆化 ^3H-TDR 信号肽 受体细胞 成肌细胞 

分 类 号：R737[医药卫生—肿瘤] R735[医药卫生—临床医学]