Experimental Study On The Reversing Effect To HepG2/ADM Cell Of Recombinant Adenovirus Vector Carring MDR1-Antisence RNA

PART ONEObjective To induce the hepatocelluar carcinoma cell(HCC) line HepG2 into multidrug resistance cell line HepG2/ADM.Mothods The agent of ADM whose dose from 0.01μg/ml to 2μg/ml was used the HCC line HepG2 into mutidrug resistance cell line HepG2/ADM, which have the 90% survival in the DMEM culture medium containing 2μg/ml ADM in end-concentration and can reproduction, regain consciousness and stored in liquid nitrogen. To abserve the HepG2 and HepG2/ADM cells antidurg activity to ADM,5-Flurouracil (5-FU),Mitomycin( MMC) and Methotrexate( MTX) at the dose of 1Cmax,10Cmax, 20Cmax,30Cmax, the MTT's analyse was used. The immunohistochemical method was also used to appraise the P-glycoprotein170 expression on the HepG2/ADM and HepG2 cell membrane , Flow cytometer was to investigate the concentration of Rhodamine-123 in the HepG2/ADM cell.Results The HepG2/ADM cell line have high efficient antidurg activity to the agents of ADM ,5-FU, MMC and MTX. The P-glycoprotein170(P-gp170) was deceted on the HepG2/ADM cell membrane with immunohistochemical method but not on HepG2 cell. Flow cytometer (FCM) show the concentration of Rhodamine-123 in HepG2/ADM was very low and there was obvious remainder of Rhodamine-123 in HepG2. Conclusion The HCC cell line HepG2 can be successfully induced into the multidrug resistance cell HepG2/ADM with step by step inducion.