Establishment of a multidrug resistance cell line of human hepatic carcinoma HepG2/ADM with adriamycin and the expression analysis of multidrug resistance-associated gene

Objective To establish a multidrug resistance cell line of human hepatic carcinoma and study the mechanism of multidrug resistance.Methods Human hepatocellular carcinoma cell line HepG2 was induced to multidrug resistance cell line HepG2/ADM by intermittent administration of high dose ADM.The multidrug resistance was detected by using MTT assay.The expressions of mdr-1 gene-coded P-glycoportein(P-gp),multidrug resistance-associated protein(MRP),and GSH / GST were examined by flow cytometric assay.The expressions of mdr-1 gene-coded P-glycoportein(P-gp),multidrug resistance-associated protein(MRP),and GSH / GST were examined by flow cytometric assay.The expression of MDR and MRP gene were also detected using RT-PCR in both HepG2/ADM and HepG2 cell lines.Results HepG2/ADM was resistant to many anti-tumor agents.The IC50 of ADM was 89.38 times higher than that of HepG2.It's expression of P-gp was increased significantly to(45.5±5.1)%,while that of HepG2 were only(11.3±1.2)%.There was no difference of the MRP and GSH/GST expressions between HepG2/ADM and HepG2 cells.The level of MDR mRNA expression was relatively high in HepG2/ADM.Conclusion HepG2/ADM was a model with multidrug resistance and the level of MDR mRNA expression were higher in HepG2/ADM than that in HepG2.