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:
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- 1. Michael WM, Robert O, Fanning E, et al. Activation of DNA replication check point though RNA synthesis by primase [J].Science, 2000; 289(22)∶2133.
- 2. 邓尚平，周文碧．A14或A19125I胰岛素的制备及其意义 [J].四川医学院学报，1981； 12(4)∶275.
- 3. 李前伟，谭天秩． 125IVIP的制备及其与SGC7901人胃癌细胞受体体外结合特性研究 [J]. 中华核医学杂志, 1998； 18(2)∶73.
- 4. 匡安仁， 周绿漪， 梁正路, 等．放射性碘标记胰岛素在荷肝癌H22小鼠体内的特趋性研究 [J]. 中华核医学杂志, 1999；19(3)∶.
- 5. Kurtaran K, Li SR, Rader M, et al. Technetium99mgalactocylneoglycoalbumin combined with iodine123Tyr(A14)insulin visualized human hepatocellular carcinomas [J]. J Nucl Med, 1995; 36(10)∶1875.
- 6. Frittilta L, Vignori R, Stampfer MR, et al. Insulin receptors over expression in 184B5 human mammary epithelial cells induced a liganddependent transformed phenotype [J]. J Cell Biochem, 1995; 57(4)∶666.
- 7. Najjar SM, Choice CV, Soni P. Effect of PP120 on receptormediated insulin endocytosis regulated by the juxtamembrane domain of the insulin receptor [J]. J Bio Chem, 1998; 273(21)∶12923.
- 8. Shah N, Zhang S, Harada S, et al. Electron microscopic visualization of insulin translocation into the cytoplasm and nuclear of intact H35 hepatoma cells using covalently linked Nanogoldinsulin [J]. Endocrinology, 1995; 136(7)∶2825.
- 9. Linde S. Monoiodoinsulin specifically substituted in Try A14 or A19 [J]. Int J Peptide Protein Res, 1980; 15(3)∶495.
- 10. Marshall S. Kinetic of insulin receptor internalistion and recycling in adipocytes; shunting of receptors to degradative pathway by inhibitors of recycling[J]. J Biochem, 1985; 117(5)∶436.
- 11. Smith RM, Jarett L. Partial characterization of mechanism of insulin accumulation in H35 hepatoma cell nuclei [J].Diabetes, 1990; 39(6)∶683.
- 12. Taouis M, Derouet M, Caffin JP, et al. Increased insulin receptor number and insulin responsiveness in a chicken hepatoma cell line [J]. Eur J Biochem, 1994; 221(3)∶1127.
- 13. 179.
