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目的克隆编码多药耐药基因(mdr1)的启动子片段并探讨其在上皮性卵巢癌中的转录活性。方法2002年9月至2003年4月华中科技大学同济医学院附属协和医院以人类基因组为模板扩增人类mdr1基因的启动子片断,并将启动子分别插入到pGEMT载体和荧光素酶报告基因载体中。将重组荧光素酶报告基因系统分别转染紫杉醇耐药(A2780/Taxol)和紫杉醇敏感细胞(A2780),比较启动子在两种细胞中的活性;同时将转染后的A2780/Taxol用曲古霉素A(TSA)处理,观察TSA对启动子转录活性的影响。结果mdr1启动子在耐药细胞A2780/Taxol中表现出比A2780强的启动子活性,这种活性在TSA诱导后能够提高。结论成功克隆了mdr1启动子片段,在A2780/Taxol细胞中mdr1启动子的功能活性的研究为下一步对多耐药性卵巢癌进行靶向基因治疗提供了重要依据。
Objective To clone a promoter fragment encoding multidrug resistance gene (mdr1) and investigate its transcriptional activity in epithelial ovarian cancer. Methods From September 2002 to April 2003, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology extended the promoter region of human mdr1 gene using human genome as a template, and inserted the promoter into pGEMT vector and luciferase reporter gene vector in. The recombinant luciferase reporter gene system was transfected with paclitaxel-resistant (A2780 / Taxol) and paclitaxel-sensitive cells (A2780) respectively, and the activity of the promoter in both cells was compared. Meanwhile, the A2780 / To investigate the effect of TSA on the promoter transcriptional activity. Results The mdr1 promoter showed stronger promoter activity in A2780 / Taxol than A2780, which could be increased after induction by TSA. Conclusion The successful cloning of mdr1 promoter and the functional activity of mdr1 promoter in A2780 / Taxol cells provide an important basis for targeted gene therapy in multidrug resistant ovarian cancer.