Objective: Recurrent ovarian cancer is often resistant to drugs such as paclitaxel. Short hairpin RNA (shRNA) targeting MDRI, a gene involved in the process of drug resistance, may be a promising strategy to overco...Objective: Recurrent ovarian cancer is often resistant to drugs such as paclitaxel. Short hairpin RNA (shRNA) targeting MDRI, a gene involved in the process of drug resistance, may be a promising strategy to overcome drug resistance. Methods: Construction and identification of eukaryotic expression plasmid was transiently transfected into human ovarian cancer ce plasmid of shRNA targeting on MDR1 gene. The ne A2780/Taxol. Apoptosis was determined by flow cytometry using annexin V-FITC/PI double labeling. Expression of MDRI mRNA was detected by quantitative polymerase chain reaction (qPCR) and P-glycoprotein expression was detected using Western blot. Results The IC50 of paclitaxel in MDR1 shRNA-transfected group was significantly reduced (1.986±0.153)μmol/ml as compared with that in negative control (5.246±0.107)μmol/ml and empty vector-transfected group (5.212±0.075)μmol/ml (P〈0.05). The percent of the relative reverse sensitivity to paclitaxel on A2780/Taxol cells was 67.1%, and the apoptotic rate was significantly increased [(6.977±0.333)%] compared with control [(1.637±0.111)%] and empty vector-transfected group [(1.663±0.114)%] (P〈0.05). Expressions of MDR1 mRNA and P-glycoprotein were significantly reduced compared with control (P〈0.05). Conclusion: The present study demonstrated that the eukaryotic expression plasmid of shRNA targeting on MDRI inhibited the expression of MDRI effectively, thus enhance the sensitivity of A2780/Taxol cells to paclitaxel.展开更多
基金support from laboratory of the second hospital and medical college of Shandong university
文摘Objective: Recurrent ovarian cancer is often resistant to drugs such as paclitaxel. Short hairpin RNA (shRNA) targeting MDRI, a gene involved in the process of drug resistance, may be a promising strategy to overcome drug resistance. Methods: Construction and identification of eukaryotic expression plasmid was transiently transfected into human ovarian cancer ce plasmid of shRNA targeting on MDR1 gene. The ne A2780/Taxol. Apoptosis was determined by flow cytometry using annexin V-FITC/PI double labeling. Expression of MDRI mRNA was detected by quantitative polymerase chain reaction (qPCR) and P-glycoprotein expression was detected using Western blot. Results The IC50 of paclitaxel in MDR1 shRNA-transfected group was significantly reduced (1.986±0.153)μmol/ml as compared with that in negative control (5.246±0.107)μmol/ml and empty vector-transfected group (5.212±0.075)μmol/ml (P〈0.05). The percent of the relative reverse sensitivity to paclitaxel on A2780/Taxol cells was 67.1%, and the apoptotic rate was significantly increased [(6.977±0.333)%] compared with control [(1.637±0.111)%] and empty vector-transfected group [(1.663±0.114)%] (P〈0.05). Expressions of MDR1 mRNA and P-glycoprotein were significantly reduced compared with control (P〈0.05). Conclusion: The present study demonstrated that the eukaryotic expression plasmid of shRNA targeting on MDRI inhibited the expression of MDRI effectively, thus enhance the sensitivity of A2780/Taxol cells to paclitaxel.