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Interfering Breast Cancer Resistance Protein(BCRP) Gene For Reversing The Multidrug Resistance Of Hepatocellular Carcinoma

Posted on:2010-08-02Degree:MasterType:Thesis
Country:ChinaCandidate:T LiFull Text:PDF
GTID:2144360278465124Subject:Surgery
Abstract/Summary:PDF Full Text Request
Objective: To investigate the suppression of BCRP gene and protein by small interfering RNA (siRNA) and the restoration of sensitivity to chemotherapeutic drugs in multidrug-resistant hepatocellular carcinoma cell line HepG2/ADM.Methods: BCRP targeted siRNA duplexes were transfected into HepG2/ADM by pSUPER. The suppression of BCRP gene and protein product was analyzed by RT-PCR and Western blot, respectively. MTT assay was performed to measure the reverse effect of BCRP-siRNA based on the results of IC50.Results: The levels of BCRP mRNA and BCRP of HepG2/ADM cells were about 8-fold and 9-fold to those of HepG2, respectively. While BCRP and BCRP mRNA of HepG2/ADM were significantly inhibited by 62.41%±2.1% and 67.8%±1.8% after transfected with BCRP-siRNA- pSUPER. The over-expression of BCRP gene was suppressed efficiently by the siRNA. The levels of BCRP mRNA and protein expression in the transfected HepG2/ADM cells reduced to 32.55%±1.8% and 37.49%±2.1% in comparsion with those in the controls, respectively(P<0.01). Their resistance to ADM was reversed by 3.55 folds, which indicated the restoration of sensitivity to drugs.Conclusion: The expression of BCRP mRNA and BCRP of HepG2/ADM cells were more than those of HepG2 respectively. The sequence specific RNAi could inhibit BCRP mRNA and protein expression in the HepG2/ADM cells. It may reverse multidrug resistance phenotype; therefore, provide promising therapeutic modalities in the treatment of hepatocellular carcinoma (HCC).
Keywords/Search Tags:multidrug resistance, BCRP, RNA interference, hepatocellular carcinoma
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