IGF-ⅠR、IGF-ⅡR反义基因对SMMC-7721肝癌细胞生物学行为的影响

Effects of IGF-ⅠR, IGF-ⅡR antisense gene transfection on the biological behaviors of SMMC-7721 human hepatoma cells

目的 研究胰岛素样生长因子Ⅰ型及Ⅱ型受体 (IGF ⅠR、IGF ⅡR)反义基因对SMMC 772 1肝癌细胞生物学行为的影响。方法 应用反义技术 ,在已转染了IGF ⅠR反义基因的肝癌SMMC 772 1 (772 1 IGF ⅠR AS)细胞基础上 ,继续利用IGF ⅡR反义基因真核表达系统 ,通过DOTAP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 772 1 IGF R AS细胞。采用光镜、电镜观察形态学变化 ,双层软琼脂克隆形成实验、MTT细胞生长曲线实验及裸鼠皮下接种实验观察转染后细胞体外生长情况。结果 772 1 IGF R AS细胞在光镜、电镜下的肿瘤细胞形态恶性程度减低 ;在软琼脂培养基中的生长能力降低 (P <0 .0 1 ) ;在裸鼠体内的致瘤能力也降低 (P <0 .0 5) ;但细胞生长曲线与对照组比较则无明显差异。结论 IGF ⅠR、IGF ⅡR反义基因对SMMC 772 1肝癌细胞株的恶性行为有明显抑制作用。利用反义技术干预肝癌细胞生长因子受体基因表达可能给肝癌的基因治疗提供新的途径。

Objective To investigate the effect of IGF ⅠR, IGF ⅡR antisense gene transfection on the biological behavior of SMMC 7721 human hepatoma cells. Methods 7721 IGF ⅠR AS cells (human hepatoma SMMC 7721 cells transfected with IGF ⅠR antisense gene) were transfected with a plasmid vector expressing IGF ⅡR cDNA in the antisense orientation by DOTAP liposome 7721 IGF R AS cells were obtained by selection with G418 and hygromycin. Morphological changes of cells were observed with optic and electron microscopes. In vitro growth of the 7721 IGF R AS cells was observed with soft agar test, MTT test and with nude mice inoculation test in vivo . Results The following changes were found in the SMMC 7721 cells after transfected with the IGF IR and IGF ⅡR antisense genes: ① The degree of malignancy of the tumor cells as revealed by cell morphology was ameliorated. ② The growth capability of SMMC 7721 cells in soft agar and tumorigenicity in nude mice were significantly decreased. However, in the control group, the SMMC 7721 cells transfected with IGF IR and IGF ⅡR sense cDNA and SMMC 7721 cells transfected without any external genes had no such changes. But, the cell growth curves had no significant differences among these three groups. Conclution IGF ⅠR and IGF ⅡR antisense gene could restrain the malignant behavior of human hepatoma cell significantly, and suggest a potential route for human tumor gene therapy.