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目的观察多药耐药基因1(mdr1)启动子调控腺病毒介导的胞嘧啶脱氨酶∷尿嘧啶磷酸核糖转移酶(CD∷UPP)融合基因系统对人卵巢癌紫杉醇耐药细胞的靶向杀伤作用。方法2004年10月至2005年5月于华中科技大学附属协和医院,将重组腺病毒(Admdr1-CD∷UPP)转染卵巢癌紫杉醇耐药细胞(SKOV3/Taxol)及非耐药细胞(SKOV3),荧光显微镜下观察转染效率,用逆转录-聚合酶链反应(RT-PCR)检测目的基因CD∷UPP的表达,并给予不同浓度的5-氟胞嘧啶(5-FC),用四甲基偶氮唑蓝(MTT)法检测二组的细胞存活率及旁观者效应。结果重组腺病毒对SKOV3/Taxol的转染率随腺病毒滴度的递增而增加,感染复数(MOI)为50时,感染率约100%;CD∷UPP基因仅在SKOV3/Taxol中稳定表达;5-FC对耐药细胞转基因组的杀伤作用显著高于非耐药细胞转基因组,前者表现出明显的旁观者效应。结论mdr1启动子调控的CD∷UPP融合自杀基因系统对人卵巢癌紫杉醇耐药细胞有靶向杀伤作用。
Objective To investigate the effect of multidrug resistance gene 1 (mdr1) promoter on the regulation of paclitaxel resistant human ovarian cancer cell line mediated by adenovirus-mediated cytosine deaminase-uracil phosphoribosyltransferase (CD :: UPP) Killing effect. METHODS: The ovarian cancer cell line SKOV3 / Taxol and SKOV3 were transfected with recombinant adenovirus (Admdr1-CD :: UPP) from October 2004 to May 2005 at the Union Hospital of Huazhong University of Science and Technology. , The transfection efficiency was observed under a fluorescence microscope, and the expression of the target gene CD :: UPP was detected by reverse transcription-polymerase chain reaction (RT-PCR). Different concentrations of 5-fluorocytosine (5-FC) The cell viability and bystander effect of two groups were detected by MTT assay. Results The transfection rate of SKOV3 / Taxol was increased with the increase of adenovirus titer. The infection rate was about 100% at the MOI of 50; the expression of CD :: UPP gene was only stably expressed in SKOV3 / Taxol; The cytotoxicity of 5-FC to drug-resistant cell transgenics was significantly higher than that of non-drug resistant cells. The former showed obvious bystander effect. Conclusion The CD :: UPP fusion suicide gene system regulated by mdr1 promoter has a targeted killing effect on paclitaxel resistant human ovarian cancer cells.