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目的 研究人胰岛素样生长因子Ⅱ型受体 (IGF ⅡR)反义基因对SMMC 772 1人肝癌细胞生长的抑制作用。方法 构建IGF ⅡR正、反义基因真核表达质粒 ,用磷酸钙 DNA共沉淀的方法 ,导入SMMC 772 1人肝癌细胞株 ,经G418培养液筛选稳定的抗性细胞。采用Northern杂交检测对转染细胞内源性IGF ⅡR基因转录的作用 ,观察转染细胞克隆形成率 ,细胞生长曲线 ,软琼脂生长能力的变化。结果 IGF ⅡR反义基因转染肝癌细胞的内源性IGF ⅡR基因转录受到有效抑制 ,转染细胞克隆形成率明显降低 ,并失去在软琼脂上生长能力 ,但细胞生长曲线无明显变化。结论 IGF ⅡR反义基因可有效阻断IGF Ⅱ至IGF ⅡR的自分泌 /旁分泌生长刺激信号环路 ,一定程度抑制肝癌细胞的恶性表型。
Objective To study the inhibitory effect of human insulin-like growth factor type I receptor (IGF IIR) antisense gene on the growth of SMMC 772 1 human hepatoma cells. Methods The eukaryotic expression plasmids of IGF IIR antisense gene were constructed and co-precipitated by calcium phosphate DNA. Then SMMC 772 1 human hepatocellular carcinoma cell line was induced and stable resistant cells were selected by G418 medium. Northern blotting was used to detect the transcription of endogenous IGF IIR gene in transfected cells. The clone formation rate, cell growth curve, and soft agar growth ability of the transfected cells were observed. Results The transcription of endogenous IGF IIR gene in hepatoma cells transfected with antisense IGF antisense gene was effectively inhibited. The colony forming rate of transfected cells was significantly decreased and the ability to grow on soft agar was lost. However, there was no significant change in cell growth curve. Conclusion IGF IIR antisense gene can effectively block the signal loop of autocrine/paracrine growth of IGF II to IGF IIR, and inhibit the malignant phenotype of hepatoma cells to some extent.