HepG_2人肝癌细胞胰岛素样生长因子1受体定量分析

Quantitative analysis of IGF1Rs on human hepatocellular HepG_2 cells

目的 对人肝癌HepG2 细胞上的胰岛素样生长因子 1受体 (type 1receptorofinsulin likegrowthfactor ,IGF1R)进行定量分析。方法 常规培养HepG2 细胞 ,以Iodogen法 13 1I标记抗IGF1R单克隆抗体 1H7,SephadexG 5 0纯化标记反应液 ,13 1I 1H7和非标记 1H7与HepG2 细胞表面的IGF1R竞争结合 ,Scatchard法分析HepG2 细胞上IGF1R的平均表达量。结果 每孔HepG2 细胞 (2 0× 10 5)表面IGF1R的Bmax为 6 746× 10 -12 mol/L ,平均单个HepG2 细胞IGF1R的密度为 2 .0 3×10 4/细胞。结论 本实验条件下制备的 13 1I 1H7满足作为IGF1R特异性配体的要求 ,碘标记 1H7对其与IGF1R结合活性无明显影响 ,HepG2 细胞表达高密度与高亲和力的IGF1R。

Objective To quantitatively analyze type 1 receptor of insulin-like growth factors (IGF1Rs) on the HepG 2 cells. Methods 1H7 (monoclonal antibody against IGF1R) was labeled with radionuclide 131 I by Iodogen method. The mixture was purified with Sephadex G-50 column, 131 I-1H7, and its unlabeled counterpart 1H7 competitively bound to IGF1Rs on the HepG 2 cells. The average number of IGF1R on the HepG 2 cells was analyzed by Scatchard method. Results The Bmax of HepG 2 cells (2.0×10 5) in every well was 6.746×10 -12 mol/L. The expression of IGF1R on the HepG 2 cells was 2.03×10 4/cell. Conclusion IGF1Rs are expressed on the HepG 2 cells with high density and high affinity, which may be used in other related studies.