摘要
AIM: To reverse multidrug resistance (MDR) of HepG2 by anti-MDR1 hammerhead ribozyme.METHODS: We developed an anti-MDR1 hammerhead ribozyme and delivered it to P-gp-overproducing human hepatocarcinoma cell line HepG2 by a retroviral vector containing RNA polymerase Ⅲ promoter. We detected the expression of mdr1/Pgp and Rz in HepG2, HepG2 multidrugresistant cell line and HepG2 Rz-transfected cells by realtime RT-PCR, semi-quantitative RT-PCR and Western blot methods. Moreover, MTT assay was tested to detect sensitivity of these ribozyme-transfected cells, and Rhodamine123 (Rh123) applied to test the function of Pgp.RESULTS: The Rz- transfected HepG2 cells became doxorubicin-sensitive, concomitant with the decreases in MDR1 expression, P-gp amounts and efflux pump function.CONCLUSION: The approaches using either retrovirus or liposome-mediated transfer of anti-MDR1 ribozyme may be selectively applicable to the treatment of MDR cells.
AIM:To reverse multidrug resistance (MDR) of HepG2 by anti-MDR1 hammerhead ribozyme. METHODS:We developed an anti-MDR1 hammerhead ribozyme and delivered it to P-gp-overproducing human hepatocarcinoma cell line HepG2 by a retroviral vector containing RNA polymerase Ⅲ promoter.We detected the expression of mdr1/Pgp and Rz in HepG2,HepG2 multidrug- resistant cell line and HepG2 Rz-transfected cells by real- time RT-PCR,semi-quantitative RT-PCR and Western blot methods.Moreover,MTT assay was tested to detect sensitivity of these ribozyme-transfected cells,and Rhodamine123 (Rh123) applied to test the function of Pgp. RESULTS:The Rz-transfected HepG2 cells became doxorubicin-sensitive,concomitant with the decreases in MDR1 expression,P-gp amounts and efflux pump function. CONCLUSION:The approaches using either retrovirus or liposome-mediated transfer of anti-MDR1 ribozyme may be selectively applicable to the treatment of MDR cells.
基金
the Clinical Focal Point Subject Foundation of Ministry of Public Health
