Establish hepatocellular carcinoma multidrug resistance cell line by transferring the MDR1 cDNA

Objective To establish a stable HCC MDR cell line on the basis of transgenic mdr1.Methods The 4.5-kb mdr1 cDNA was transferred into human hepatocarcinoma cell line HepG2 by the PCI-neo mammalian expression vector and the transfected HepG2 cells resisting G418 were proliferated.Then the mdr1 mRNA and P-gp in these HepG2 cells were detected by the means of RT-PCR and FCM respectively.The accumulation of the daunorubicin was determined by FCM simultaneously.The nude mice model of grafting tumor was established by injecting subcutaneously HepG2/mdr1 cells in the right axilla and ADM was injected into peritoneal cave when the tumor diameter reached 5mm.The size and growth inhibition of the tumor were evaluated.Results The MDR hepatocarcinoma cell line HepG2/mdr1 was developed and the mdr1 mRNA and P-gp in HepG2/mdr1 cells were(58.80±11.80)%and(10.28±2.09)respectively,compared with(24.00±5.80)% and(3.70±1.06)in HepG2 cells.The resistance to ADM and MMC increased 35 and 125 times respectively.In the nude mice HCC model,after ADM therapy,the mean size of HepG2 cell tumors was obviously smaller than HepG2/mdr1 cell tumors,the difference was significant.Conclusions The approach using the transfer of mdr1 cDNA may be applicable to the development of MDR hepatocarcinoma cell line which would provide the experimental basis of MDR research.