腺病毒介导CD/UPRT自杀基因系统治疗胰腺癌的实验研究  

Gene therapy for pancreatic cancer by using adenovirus-mediated transfer of cytosine deaminase/uracil phosphoribosyltransferase

作　　者：张世能[1] 徐凤琴[1] 于钟[1] 左海军[1] 危淑莉[1] 黄志清[1] 

机构地区：[1]中山大学附属第二医院消化内科,广州510120

出　　处：《胰腺病学》2007年第6期372-374,共3页Chinese JOurnal of Pancreatology

基　　金：广东省自然科学基金（04009381）

摘　　要：目的探讨CD/UPRT自杀基因系统在胰腺癌基因治疗中的作用。方法采用同源重组技术构建重组腺病毒Ad-CD、Ad-CD/UPRT和Ad-GFP;体外感染人胰腺癌SW1990细胞,RT-PCR检测UPRTmRNA表达;MTT法检测细胞对5-FC的敏感性及旁观者效应;流式细胞仪分析细胞凋亡率;建立胰腺癌裸鼠皮下移植瘤模型,瘤体内直接注射腺病毒,观察CD/UPRT自杀基因的原位治疗作用。结果转染CD/UPRT的胰腺癌SW1990细胞对5-FC的敏感性明显提高,Ad-CD/UPRT组的IC50为25μmol/L,而Ad-CD组和Ad-GFP组分别为100μmol/L和〉1000μmol/L。Ad-CD组、Ad-CD/UPRT组和Ad-GFP组的细胞凋亡率分别为61.3%、40.5%和3.0%,3组比较有显著差异（P〈0.05）。CD/UPRT基因转染存在旁观者效应。Ad-CD/UPRT瘤内注射治疗后种植癌体积缩小81%,Ad-GFP治疗肿瘤缩小63%。结论CD/UPRT自杀基因系统能提高对胰腺癌细胞的杀伤作用。Objective To investigate the effect of a combined suicide gene system of cytosine deaminase/uracil phosphoribosyltransferase on gene therapy of pancreatic cancer.Methods The recombinant adenovirus containing CD,CD/UPRT,and GFP was constructed by homologous recombinant.A human pancreatic cancer cell line SW1990 was used to detect the transfer efficiency in vitro.The expression of CD/UPRT mRNA was determined by RT-PCR.The sensitivity of SW1990 cell towards 5-FC and bystander effect was measured by MTT assay.The cell apoptosis was analyzed by flow cytometry.The growth of tumors which had been generated from SW1990 cells inoculated into nude mice was to detect the effect of the recombinant adenovirus in vivo.Results Transinfection with Ad-CD/UPRT resulted in a significant increase of sensitivity towards 5-FC compared with Ad-CD or Ad-GFP.The IC50 was 25μmol/L,100μmol/L,and 〉 1 000 μmol/L,respectively.The apoptosis cells rates were 61.3%,40.5%,and 3.0%,respectively（P 〈 0.05）.The bystander effect was observed in SW1990 cells transinfected with Ad-CD/UPRT.The anticancer effect was showed in vivo SW1990 xenografts nude mice with in situ Ad-CD/UPRT gene transduction,which was superior to Ad-CD and Ad-GFP.Conclusions The results suggested that CD/UPRT gene therapy may be a new approach in the treatment of pancreatic cancer.

关 键 词：胰腺肿瘤 胞嘧啶脱氨酶 尿嘧啶磷酸核糖转移酶 腺病毒 基因治疗 

分 类 号：R735[医药卫生—肿瘤]