A14^(125)I-胰岛素与H_(22)肝癌细胞胰岛素受体结合的研究  

The in Vitro Study of A14^(125)I-Insulin Combining with Receptor on Rat H_(22) Hepatocarcinoma Cell Membrane

作　　者：程中[1] 欧晓红[2] 匡安仁[2] 

机构地区：[1]四川大学华西医院普外科,成都610041 [2]四川大学华西医院核医学科

出　　处：《中国普外基础与临床杂志》2002年第3期168-171,共4页Chinese Journal of Bases and Clinics In General Surgery

基　　金：国家自然科学基金 (39770 2 2 9)

摘　　要：目的　体外分析H2 2 肝癌细胞与正常肝细胞膜的胰岛素受体表达 ,探讨胰岛素作为受体介导靶向治疗载体的可行性及临床应用前景。方法　用氯氨T法制备A14 12 5I 胰岛素 ,电泳分离纯化标记物并用自身置换比放射性测定和过量受体结合实验法鉴定标记物质量 ,再制备H2 2 肝癌细胞及正常小鼠肝细胞 ,进行体外受体结合实验 ,分别测定12 5I 胰岛素与小鼠H2 2 肝癌及正常肝细胞膜胰岛素受体结合的Kd值及Bmax值以及其竞争结合曲线。12 5I 胰岛素受体结合的Kd值及Bmax值用t检验分析 ,饱和结合曲线用Scatchard分析。结果　小鼠H2 2 肝癌细胞及正常小鼠肝细胞胰岛素受体Bmax值分别为 (5 .6± 1.1) pmol/10 6细胞和 (3 .2± 0 .8) pmol/10 6细胞 ,高亲和力Kd值为 (1.8± 0 .6)nmol/L及 (2 .1± 0 .9)nmol/L ,低亲和力Kd值为 (3 2 .0± 10 .7)nmol/L及 (3 7.0± 12 .3 )nmol/L。癌细胞胰岛素受体Bmax值明显高于正常肝细胞 (P<0 .0 5 )。受体结合实验表明 ,12 5I 胰岛素与H2 2 肝癌及正常细胞的结合具有特异性。结论　12 5I 胰岛素与H2 2 肝癌及正常小鼠肝细胞膜受体的结合具有高度特异性 ,H2 2 肝癌细胞较正常小鼠肝细胞表达胰岛素受体增加 ,提示胰岛素可作为抗癌载体通过与肝癌细胞表面受体结合而介导肝癌的靶向治疗。Objective To investigate the insulin receptor expression and binding characteristics of H 22 rat hepatic cancer cell membrane with 125 I insulin and the possibility of using insulin as a carrier for the receptor mediated targeted therapy. Methods Insulin was radio labelled using Ch T method, isolated and purified by polyacrylamide gel electrophoresis, the labelled 125 I insulin was measured by specific activity self replacement and over dose receptor combination experiment. The rat H 22 hepatocarcinoma cells, normal rat hepatic cells were made, continuing the value Kd and Bmax of 125 I insulin binding with insulin receptor on rat H 22 hepatocarcinoma and normal hepatic cells membrane were evaluated in vitro, the competed binding curve was pictured. The binding Kd and Bmax value of 125 I insulin receptor were evaluated with t test and receptor binding curve was tested with Scatchard method. Results The Bmax value of rat H 22 hepatocarcinoma cell receptor [(5.6± 1.1) pmol/10 6 cell] was higher than that of normal hepatic cell [(3.2±0.8) pmol/10 6 cell] significantly ( P <0.05). The Kd value of H 22 cell [high and low affinity vs (1.8±0.6) nmol/L and (32.0±10.7) nmol/L] and normal hepatic cell [high and low affinity vs (2.1±0.9) nmol/L and (37.0±12.3) nmol/L] were not significant respectively. In this experiment, it was specific when 125 I insulin combined with receptor on H 22 hepatocarcinoma cell and rat normal hepatic cell membrane. Conclusion This experiment showed that the receptor expresses much more significantly on H 22 hepatocarcinoma cell membrane than that on normal rat hepatic cell membrane, 125 I insulin combining with receptor on H 22 hepatocarcinoma cell and rat hepatic cell membrane is highly specific. We may use insulin as an anticancer carrier to mediate insulin combined with receptor on hepatocarcinoma cell membrane in the target treatment of hepatic cancer.

关 键 词：胰岛素 受体 肝癌 癌细胞胰岛素受体 动物实验 

分 类 号：R735.7[医药卫生—肿瘤] R73－361[医药卫生—临床医学]