mdr1启动子调控双自杀基因对卵巢癌SKOV3耐药细胞的靶向杀伤作用  

The selective killing effects of adenovirus mediated double suicide gene system controlled by MDR1 promoter on SKOV3/Taxol ovarian cancer cells

作　　者：卢实[1] 王晓翊[1] 肖蓝[1] 蔡俐琼[1] 王泽华[1] 

机构地区：[1]华中科技大学附属协和医院妇产科,湖北武汉430022

出　　处：《中国实用妇科与产科杂志》2007年第10期763-765,共3页Chinese Journal of Practical Gynecology and Obstetrics

基　　金：国家自然科学基金(30070786)

摘　　要：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 evaluate the selective killing effects of adenovirus mediated CD ：： UPP suicide gene controlled by mdrl promoter on SKOV3/Taxol ovarian cancer in vitro. Methods From October 2004 to May 2005 in the Union Hospital of Huazhong University of Science and Technology, the drug resistant SKOV3/Taxol cells and non - resistant SKOV3 cells were infected with recombined adenovirus, and the infection rate was measured by observing the expression of green fluorescent protein under the fluorescent microscopy. The expressing of CD ：： UPP gene was detected by RT - PCR method. Then 5 -fluorocytosine （5 -FC） was given in different concentrations to the infected cells ,and the relative survival rate and bystander effect were measured by MTT assay. Results The infection rate of SKOV3/Taxol cells was increased gradually with the increasing MOI of adenovirus. The infection rate was almost 100% when MOI was 50. The CD ：： UPP gene was detected in SKOV3/Taxol cells only. The killing effects of SKOV3/Taxol cells infected with Admdrl -CD ：： UPP and 5- FC were significantly higher than that of SKOV3 group （ P 〈 0.05 ）. In addition, a profound bystander effect was observed by MTT method. Conclusion The CD ：： UPP fusion suicide gene driven by mdrl promoter can confer selectively expression and killing effects on SKOV3/Taxol cells.

关 键 词：多药耐药 胞嘧啶脱氨酶 尿嘧啶磷酸核糖转移酶基因 基因治疗 卵巢癌 

分 类 号：R71[医药卫生—妇产科学]