四川大学学报（医学版）2009，Vol.40Issue(6)：983-987,5.
靶向性双自杀基因治疗载体pcDNA3.1(-)Cp-CD-TK构建的初步研究
A Preliminary Study on Construction of Targeting Double Suicide Gene Therapy Vector pcDNA3.1(-)Cp-CD-TK
摘要
Abstract
Objective To construct the targeting double suicide gene therapy vector pcDNA3. 1( -) Cp-CD-TK driven by carcino-embryonic antigen promoter (CEA promoter,Cp), and to investigate whether the vector could control the expression of CD-TK gene specificity and impact on proliferation of CEA positive colon cancer cells. Methods Three kinds of target gene, Cp, CD, and TK were obtained by PCR, the products were double digested and inserted into pcDNA3. 1(-), then the targeting vector pcDNA3. 1 (-) Cp-CD-TK was transfected into CEA positive human colon cancer SW480 cells and CEA negative Hela cells, the expression of CD-TK was examined by RT-PCR. The sensitivities of SW480 cells transfected with pcDNA3. 1 (-) Cp-CD-TK to pro-drug 5-Fluorocytosine (5-Fc) and Ganciclovir (GCV) were detected by MTT analysis. Results Recombinants with Cp, CD and TK insert were obtained. Constructed targeting gene therapy vector pcDNA3. 1 (-) Cp-CD-TK was confirmed by gel electrophoresis and sequencing. RT-PCR analysis further confirmed CD-TK gene was expressed in SW480 cells and was not expressed in CEA negative Hela cells. MTT assay demonstrated that SW480 cells transfected with targeting vector pcDNA3.1(-)Cp-CD-TK were sensitive to pro-drug 5-Fc and GCV. Conclusion Targeting double suicide gene therapy vector pcDNA3. 1 (- ) Cp-CD-TK was constructed correctly. The vector could make CD-TK gene express specifically in CEA positive cells for the purpose of targeting killing colon cancer.关键词
双自杀基因/癌胚抗原基因启动子/CD/TK/大肠癌Key words
Suicide gene/CEA promoter /CD /TK /Colorectal carcinoma分类
医药卫生引用本文复制引用
罗琪,熊宇,卢毅卓..靶向性双自杀基因治疗载体pcDNA3.1(-)Cp-CD-TK构建的初步研究[J].四川大学学报（医学版）,2009,40(6):983-987,5.基金项目
厦门市卫生局重点项目基金(WSK0509)资助 （WSK0509）
