酶受体分析法测定3AO细胞LH/hCG受体结合特性  

Characterization of LH/hCG Receptor by Enzyme Receptor Assay in Human Ovarian Cancer Cell Line 3AO

作　　者：邢艳[1] 廖英[2] 陈曼玲[2] 

机构地区：[1]川北医学院附属医院检验科,四川南充637000 [2]四川大学华西医学中心基础与法医医学院,四川成都610041

出　　处：《标记免疫分析与临床》2005年第1期27-31,共5页Labeled Immunoassays and Clinical Medicine

摘　　要：建立培养细胞的酶受体分析法(enzyme receptor assay, ERA),应用ERA研究卵巢癌细胞株3AO的LH/hCG受体特性。采用过碘酸钠法制备辣根过氧化物酶(horseradish peroxidase, HRP) 人绒毛膜促性腺激素(human chorionic gonadotropin, hCG)的酶标配体交联物, 与培养包被的3AO细胞在适当的条件下建立 ERA,并以人卵巢癌细胞株SKOV 3作为阴性对照, 测定3AO细胞膜上LH/hCG受体的竞争结合、特异性结合、饱和结合特性并计算受体的最大结合容量(Bmax)和解离常数(Kd)。ERA测得 3AO细胞 LH/hCG受体的 Bmax为 3.65mmol/g蛋白, Kd 为0.41nmol/L,未标记配体 hCG抑制 LH/hCG受体对酶标配体(HRP hCG)特异结合的IC50为0.92μmol/L。结论: 竞争结合实验、特异性结合实验、饱和结合实验表明 3AO细胞膜上存在高亲和力的LH/hCG受体。To establish an enzyme receptor assay (ERA) for LH/hCG receptor on intact culture cells and study the characteristics of LH/hCG receptor of ovarian cancer cell line 3AO by ERA, the enzyme-ligand conjugate was prepared by the oxidation method of NaIO 4. An ERA of marker ligand with embedded cells was established in suitable conditions, and applied to determine the maximum binding and affinity of LH/hCG receptor on 3AO cells as well as on an ovarian cancer cell line SKOV-3 serving as the negative control.The results showed that the B max and K d values of LH/hCG receptor were 3.65mmol/g protein and 0.41nmol/L, respectively.For competitive inhibition, the IC 50 of unlabelled hCG against HRP-hCG binding was 0.92μmol/L.Our conclusion is that by competitive inhibition assay, saturation binding assay and Scatchard analysis, LH/hCG receptors with high affinity are present on 3AO cells.

关 键 词：卵巢肿瘤 人绒毛膜促性腺激素 LH/hcG受体 酶受体分析法 

分 类 号：R737.31[医药卫生—肿瘤]