磷酸钙纳米介导自杀基因载体的构建及其应用研究(英文)  

Construction of Calcium Phosphate Nanoparticles Mediating Fusion Suicide Gene yCDglyTK and its Application in vitro

作　　者：张利元[1] 刘霆[1,2] 颜金鹏[1] 杨魁[2] 余术宜[1] 颜世能[1] 谢薇[1] 陈玉祥[1] 

机构地区：[1]中南大学生物科学与技术学院,中国湖南长沙410013 [2]中南大学湘雅医院,中国湖南长沙410008

出　　处：《生命科学研究》2009年第2期109-115,共7页Life Science Research

基　　金：Foundation item:95national new drug research projec(t96-901-05-138)~~

摘　　要：构建由CEA启动子、CMV增强子驱动的融合自杀基因PCDNA3.1(-)CVyCDglyTK表达载体和分别由CEA启动子和巨细胞病毒(CMV)增强子驱动的融合自杀基因PCDNA3.1(-)CEAyCDglyTK、PCDNA3.1(-)CMVyCDglyTK表达载体,以磷酸钙纳米为载体分别转染CEA阳性的人结肠癌细胞株LOVO细胞和CEA阴性的HeLa细胞,Lovo细胞在感染以上3种质粒表达载体后均有yCDglyTK mRNA表达,且对5-FC的敏感性明显增强;HeLa细胞在纳米PCDNA3.1(-)CMVyCDglyTK复合物感染后有yCDglyTK mRNA表达,对5-FC的敏感性增强,而在另外两种复合物感染后则没有yCDglyTK mRNA表达,5-FC对其亦无杀伤作用.结果表明靶向性基因治疗载体能使融合自杀基因在CEA阳性细胞中专一性表达,达到靶向治疗肿瘤的目的.The three expressing plasmids of pcDNA3.1 （-）CVyCDglyTK,pcDNA3.1 （-）CEAyCDglyTK, pcDNA3.1 （-）CMV-yCDglyTK were constructed with carcino-embryonic antigen （CEA） promoter and cytomegalovirus （CMV）, CEA promoter, cytomegalovirus （CMV） enhancer, respectively. Calcium phosphate nanoparticles （CPNP） was used as vector to transfer the three plasmids into CEA-positive cells （Lovo） and CEA-negative cells （HeLa）, respectively. The results showed that expression of yCDglyTK mRNA in all cells transferred by the three plasmids above,which significantly sensitized the cytotoxicity of prodrug 5-FC; expression of yCDglyTK mRNA in HeLa cells after transfected by CPNP- pCDNA3.1 （-）CMVyCDglyTK, which enhanced sensitivity of 5-FC,but the other had no killing effect, pcDNA3.1 （-）CVyCDglyTK gene therapy. plasmids had no expression of yCDglyTK mRNA might be a promising candidate vector for colon and 5-FC carcinoma gene therapy.

关 键 词：癌胚抗原 纳米载体 自杀基因 基因治疗 

分 类 号：Q782[生物学—分子生物学]