Objective To investigate the insulin receptor expression and binding characteristics of H22 rat hepatic cancer cell membrane with 125Iinsulin and the possibility of using insulin as a carrier for the receptor mediated targeted therapy. MethodsInsulin was radiolabelled using ChT method, isolated and purified by polyacrylamide gel electrophoresis, the labelled 125Iinsulin was measured by specific activity selfreplacement and over dose receptor combination experiment. The rat H22 hepatocarcinoma cells, normal rat hepatic cells were made, continuing the value Kd and Bmax of 125Iinsulin binding with insulin receptor on rat H22 hepatocarcinoma and normal hepatic cells membrane were evaluated in vitro, the competed binding curve was pictured. The binding Kd and Bmax value of 125Iinsulin receptor were evaluated with t test and receptor binding curve was tested with Scatchard method. ResultsThe Bmax value of rat H22 hepatocarcinoma cell receptor [(5.6±1.1) pmol/106 cell] was higher than that of normal hepatic cell [(3.2±0.8) pmol/106 cell] significantly (P<0.05). The Kd value of H22 cell [high and lowaffinity 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 125Iinsulin combined with receptor on H22 hepatocarcinoma cell and rat normal hepatic cell membrane. Conclusion This experiment showed that the receptor expresses much more significantly on H22 hepatocarcinoma cell membrane than that on normal rat hepatic cell membrane, 125Iinsulin combining with receptor on H22 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.
Citation: CHENG Zhong,OU Xiaohong,KUANG Anren. The in Vitro Study of A14125IInsulin Combining with Receptor on Rat H22 Hepatocarcinoma Cell Membrane. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2002, 9(3): 168-171. doi: Copy