IGF-Ⅰ及IGF-Ⅱ受体反义基因对SMMC-7721肝癌细胞株增殖能力的影响

Effect on proliferation capability of SMMC-7721 human hepatoma cell strain by IGF-ⅠR?IGF-ⅡR antisense gene transfection

目的 研究胰岛素样生长因子Ⅰ受体、Ⅱ受体 (IGF ⅠR、IGF ⅡR)反义基因对SMMC 772 1细胞体外增殖能力的影响。方法 应用反义技术 ,在已将IGF ⅠR反义基因转染入SMMC 772 1肝癌细胞株的基础上 (获得ⅠR AS细胞 ) ,继续利用IGF ⅡR反义基因真核表达系统 ,通过DOTAPTM脂质体介导的转染技术 ,将IGF ⅡR反义基因导入该细胞 ,经G41 8及潮霉素双重筛选获得稳定表达IGF ⅠR、IGF ⅡR反义基因的SMMC 772 1细胞 (Ⅰ、ⅡR AS细胞 )。结果 光镜、电镜下反义转染细胞形态学上肿瘤性有一定减弱 ;在软琼脂培养基中生长能力低下 (P <0 0 1 ) ;反义G41 8抗性转染细胞克隆形成能力低下 (P <0 0 5) ;在细胞生长曲线上与对照组相比有轻度下降趋势。结论 IGF ⅠR、IGF ⅡR反义基因对SMMC 772 1肝癌细胞株的恶性增殖有明显抑制作用。

Objective To investgate the effect on proliferation capability of SMMC 7721 human hepatoma cell by IGF Ⅰ?IGF ⅡR antisense gene.Methods On the base of ⅠR AS cell(we have transfected human hepatoma cell line SMMC 7721 with IGF ⅠR antisense gene), continuely, we transfected ⅠR AS cell with a plasmid vector expressing IGF ⅡR cDNA in the antisense orientation using DOTAP TM liposome. the result of PCR exhibited that the the recombinant containing IGF ⅡR antisense gene have integrated into chromosome of ⅠR AS cell.Results We observed some cell morphological changes of transfected cells by light microscope and electronical microscope;the growth capability in soft agar obviously decreased; antisense transfection clone with G418 resistance was decreased too;the growth curve of antisense transfected cells has gently descent tendency compare with control groups.Conclusion IGF ⅠR and IGF ⅡR antisense gene could significantly restrain the malignant proliferation of human hepatoma cell