Compared Biological Characteristics Of Three Multidrug Resistant Cell Line Models Of Human Hepatic Carcinoma Bel-7402/ADM

ObjectiveTo compare biological characteristics of three types of human hepatic carcinoma multi- drug resistant cell line models Bel-7402/ADM established by vitro concentration gradient increased progressively induction, nude mice liver implanted and subcutaneous implanted induction.MethodsEstablished human hepatic carcinoma adriamycin multi-drug resistant cell line models Bel-7402/ADMV,Bel-7402/ADML and Bel-7402/ADMS by three methods of vitro concentration gradient increased progressively induction, nude mice liver implanted and subcutaneous implanted induction respectively. phase contrast micro- scope was used to observed the shape of cells, MTT assay was used to evaluate several kinds of cells drugs sensitivity, growth curves were drawn by cell counting and doubling time of cell lines were calculated according to formula, flow cytometric assay was used to examine cells adriamycin influx and efflux and the expression of p-glycoprotein(P-gp), multi- drug resistance protein (MRP) and glutathione S-transferase (GSH/GST).Results1. Cells morphology observation. Under the phase contrast microscope, growth speed about three kinds of multi-drug resistant cells reduced, the shape were not different obviously, all were irregular, the volume was bigger slightly than the parental genera- tion, the cell nucleus shrinked lightly, nude mice implanted resistant cells shape closer to the parental generation. 2. Cells sensitivity on anticarcinogens. Three groups of cells were resistant to ADM and CDDP, compared with the parental generation, IC50 were significant difference (P< 0.01), and the IC50 of Bel- 7402/ADMV was the highest, so it had greater resistance. All the cells were not resistant to MMC, MTX and 5-Fu.3. growth curve and doubling time. The doubling time of resistant cells were extended obviously to the parental generation, Bel-7402, Bel-7402/ADMS, Bel-7402/ADML and Bel-7402/ADMV doubling time were: 39h, 45h, 46h and 65h.4. Cells ADM influx and efflux. in the same conditions, the Bel-7402/ADMV had max- imum ADM efflux rate, Bel-7402/ADML and Bel-7402/ADMS were Secondly, the par- ental generation was minimum. The ADM discharge rate of four cells were: 81.06%, 66.56%, 61.56% and% 34.14%.5. Flow cytometry determine the expression of P-gp, MRP and GSH/GST. Compared with the parental generation, three kinds of multidrug resistant cells P-gp,MRP were significantly improve (P<0.01), and the expression of GSH/GST without obvious changes (P>0.05).Conclusions1. Three multidrug resistant cell line models of human hepatic carcinoma Bel-7402/ ADM established by three different methods separately have stable resistance.2. The multidrug resistant cell model established by vitro concentration gradient increased progressively induction had higher resistance index and stability, but the disadvantage was that the proliferation ability is lower, induced-process is time-consuming, and easily lead to pollution.3. The resistance index of the multidrug resistant cell lines established by nude mice liver implanted and subcutaneous implanted induction were lower slightly, its advantage was the time-expended is shorter, the chance of pollution is smaller, the proliferation ability is stronger, so this two cell lines closer to clinical drug resistance, the nude mice liver implanted one highly simulated human hepatocarcinoma especially, it was a better model because of approximating human hepatic carcinoma biology and anticancer pharmacokinetics characteristics.