EMMPRIN Mediates Multidrug Resistance In Hepatocellular Carcinoma 7721

Objective: To establish human multidrug-resistant hepatocellular carcinoma cell line in twodifferent methods. silencing EMMPRIN expression to evaluate their characteristics. Investigatethe role of EMMPRIN on multidrug-resistantce in hepatocellular carcinoma cells.Methods:1.Two independent multidrug-resistant sublines, 7721/Adm1 and 7721/Adm2,were developedfrom the established human hepatocellular carcinoma cell line SMMC-7721. 7721/Adm1 wasdeveloped by a long-term continuous exposure of 7721cells to Adriamycin in stepwiseincrements of concentration; 7721/Adm2 was selected/induced by pulse treatments by using asinge concentration of Adramycin.2.EMMPRIN specific siRNA was transfected into 7721/Adm1 and 7721/Adm2 cells. RNAi assaywas performed to silence EMMPRIN expression. Established EMMPRIN silencing cell lines7721/Adm1/RNAi and 7721/Adm2/RNAi.3.EMMPRIN,MDR-1 mRNA expression were determined with reverse transcription polymerasechain reaction(RT-PCR).4.Western blot was performed to assess the expression of EMMPRIN,P-glycoprotein(P-gp)andGAPDH.5The multidrug resistance to multianticancer agents was evaluated by MTT assay.Results: The two MDR sublines may serve as two different modles for the study of multidrugresistance in human hepatocellular carcinoma.The results show that EMMPRIN expression wasincreased in 7721/Adm1 and 7721/Adm2,as compared to 7721 cells.The MDR cells producedmore MDR-1,which increased resistance to chemotherapeutic drugs.On the otherhand,EMMPRIN silencing in 7721/Adm1 and 7721/Adm2 cells by RNAi led to the oppositeeffect.Conclusion:1.Two MDR cell line models of human hepatocellular carcinoma (7721/Adm1 and 7721/Adm2)established by two different methods separately have stable resistance.2.EMMPRIN is involved in MDR and functionally mediates tumor cells MDR in humanhepatocellula carcinoma cell line SMMC-7721.The MDR of SMMC-7721 was reversed at somedigree by EMMPRIN specific siRNA transfection.