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The in Vitro Study of A14125IInsulin Combining with Receptor on Rat H22 Hepatocarcinoma Cell Membrane

Objective To investigate the insulin receptor expression and binding characteristics of H22 rat hepatic cancer cell membrane with 125Iinsulin and the possibility of using insulin as a carrier for the receptor mediated targeted therapy. MethodsInsulin was radiolabelled using ChT method, isolated and purified by polyacrylamide gel electrophoresis, the labelled 125Iinsulin was measured by specific activity selfreplacement and over dose receptor combination experiment. The rat H22 hepatocarcinoma cells, normal rat hepatic cells were made, continuing the value Kd and Bmax of 125Iinsulin 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 125Iinsulin 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 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 125Iinsulin 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, 125Iinsulin 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.