mdr1启动子调控CD∷UPP基因对紫杉醇耐药卵巢癌细胞的杀伤作用  

Cell-killing effects of adenovirus-mediated transfer of CD∷UPP gene directed by mdr1 promoter on Taxol-resis-tant ovarian cancer cells

作　　者：卢实[1] 蔡俐琼[1] 王晓翊[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,武汉430022

出　　处：《肿瘤》2007年第7期515-517,共3页Tumor

基　　金：国家自然科学基金资助项目(编号:30070786)

摘　　要：目的:探讨腺病毒介导的mdr1启动子调控胞嘧啶脱氨酶∷尿嘧啶磷酸核糖转移酶(CD∷UPP)融合基因联合5-氟胞嘧啶(5-FC)对紫杉醇耐药卵巢癌细胞的特异性杀伤作用。方法:扩增、纯化含有mdr1-CD∷UPP基因的重组腺病毒,转染人卵巢癌紫杉醇耐药细胞株A2780/Taxol和亲本细胞株A2780,RT-PCR检测mdr1和CD∷UPP基因的表达水平;之后加入5-FC,MTT法检测细胞抑制情况及旁观者效应,并观察腺病毒转染后裸鼠移植瘤的生长情况。结果:mdr1和CD∷UPP基因在A2780/Taxol细胞中可稳定表达,转染后A2780/Taxol组的细胞生长明显低于A2780组;转基因的A2780/Taxol细胞联合5-FC后可通过旁观者效应杀伤周围未转基因的耐药细胞;耐药组移植瘤生长明显受到抑制,肿瘤体积为(569.10±187.93)mm3,对照组肿瘤体积为(2111.98±230.82)mm3,差异有统计学意义(P<0.01)。结论:mdr1启动子可调控CD∷UPP基因特异性表达并特异性杀伤紫杉醇耐药卵巢癌细胞。Objective ： To evaluate the specific killing effects of the adenovirus-mediated transfer of cytosine deaminase gene and uracil phosphoribosyltransferase gene （CD： ：UPP） directed by the mdr1 promoter combined with 5-fluorocytosine （5-FC） on Taxol-resistant ovarian cancer cells in vitro and in vivo. Methods： The adenovirus vector carrying the mdr1 -CD ：： UPP gene was amplified, purified, and transferred into Taxol-resistant A2780/Taxol ovarian cancer cells and A2780 cells. The expression of mdr1 and CD ：： UPP was detected by RT-PCR. After addition of 5-FC, the cell proliferation was assessed by MTT assay. The female BALB/c nude mice bearing tumor xenografts were injected adenovirus vector containing mdr1-CD ：： UPP gene and then administered 5-FC. The growth of transplanted tumors was observed. Results. Mdrl and CD ：： UPP gene were stably expressed in A2780/Taxol cells. The growth speed of A2780/ Taxol cells was significantly slower than A2780 cells after transfection. Transfection of CD ：： UPP gene combined with administration of 5-FC induced bystander effect of A2780/Taxol cells on neighbor cells. The tumor growth in test group was significantly inhibited compared with control group [ （569.10±187.93 ） mm^3 vs （2 111.98±230.82） mm^3, P 〈0.01 ]. Conclusion： The mdr1 promoter in an adenovirus vector regulates the specific expression of CD ：： UPP in A2780/Taxol cells. This system specifically kills Taxol-resistant ovarian cancer cells.

关 键 词：卵巢肿瘤 基因疗法 基因 mdr 基因 转基因 自杀 胞嘧啶脱氨酶 尿嘧啶磷酸核糖转移酶 

分 类 号：R737.31[医药卫生—肿瘤]