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AIM:To reverse multidrug resistance (MDR) of HepG2 byanti-MDR1 hammerhead ribozyme.METHODS:We developed an anti-MDR1 hammerheadribozyme and delivered it to P-gp-overproducing humanhepatocarcinoma cell line HepG2 by a retroviral vectorcontaining RNA polymerase Ⅲ promoter.We detected theexpression 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 blotmethods.Moreover,MTT assay was tested to detectsensitivity of these ribozyme-transfected cells,andRhodamine123 (Rh123) applied to test the function of Pgp.RESULTS:The Rz-transfected HepG2 cells becamedoxorubicin-sensitive,concomitant with the decreases inMDR1 expression,P-gp amounts and efflux pump function.CONCLUSION:The approaches using either retrovirus orliposome-mediated transfer of anti-MDR1 ribozyme may beselectively applicable to the treatment of MDR cells.
AIM: To reverse multidrug resistance (MDR) of HepG2 byanti-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 vectorcontaining RNA polymerase III promoter. theexpression 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 detectsensitivity of these ribozyme-transfected cells, and Rhodamine 123 (Rh123) applied to test the function of Pgp. RESULTS: The Rz-transfected HepG2 cells became doroxorubicin-sensitive, concomitant with the decrease in MDR1 expression, P-gp amounts and efflux pump function. either retrovirus orliposome-mediated transfer of anti-MDR1 ribozyme may beselectively applicable to the treatment of MDR cells.