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目的 :探讨核酶对膀胱癌多药耐药性的逆转效应。方法 :设计针对mdr1mRNA1959位GUC的“锤头状”(Hammerhead)核酶 (Ribozyme1 ,RZ1)基因 ,定点克隆于逆转录病毒载体 pLXSN的BamHI和EcoRI位点上 ,重组出带有核酶基因的逆转录病毒质粒pLXSN RZ1。应用脂质体DoTap将pLXSN RZ1导入到耐药的人膀胱癌细胞系BHA 60细胞中。结果 :导入pLXSN RZ1的细胞系耐药性显著降低 ,RT PCR结果显示 ,细胞内mdr1mRNA显著减少。结论 :本研究中设计的核酶能明显逆转膀胱癌细胞系多药耐药性。
Objective: To investigate the reversal effects of ribozyme on multidrug resistance in bladder cancer. Methods: The Rhamnhead ribozyme (RZZ1) gene targeting GUC at position 1959 of mdr1 mRNA was designed and cloned into the BamHI and EcoRI sites of the retroviral vector pLXSN. The reverse transcripts with ribozyme gene Vaccine plasmid pLXSN RZ1. PLXSN RZ1 was introduced into drug-resistant human bladder cancer cell line BHA 60 cells using liposomal DoTap. Results: The drug resistance of the cell lines transfected with pLXSN RZ1 was significantly reduced. RT PCR results showed that mdr1 mRNA was significantly decreased in cells. Conclusion: The ribozyme designed in this study can obviously reverse the multidrug resistance of bladder cancer cell lines.