人钠/碘转运体基因转染结肠癌SW480细胞及相关蛋白表达的检测  

Expression of human sodium/iodide symporter protein in hNIS-transfected colon cancer SW480 cells

作　　者：周佰林[1] 张琦[1] 张红征[1] 李焕斌[1] 文正伟[1] 王玲[1] 

机构地区：[1]温州医学院核医学教研室,325027

出　　处：《浙江医学》2013年第12期1120-1122,1126,共4页Zhejiang Medical Journal

基　　金：温州市科技局科研基金资助项目(Y20090094)

摘　　要：目的以重组人钠/碘转运体(hNIS)基因转染结肠癌SW480细胞并检测hNIS mRNA及蛋白的表达,为放射性碘治疗非甲状腺肿瘤提供新思路。方法将构建好的重组质粒(pcDNA3.1^+-hNIS)进行酶切、测序鉴定并扩增、提取。SW480细胞分为重组质粒(pcDNA3.1^+-hNIS)转染组、空白质粒(pcDNA3.1^+)转染组、空白对照组,转染后以RT-PCR和Western blot检测各组细胞hNIS mRNA和蛋白的表达。结果酶切和测序结果显示插入的hNIS基因大小和方向均正确。RT-PCR和Western blot显示重组质粒转染组SW480细胞可见hNIS mRNA和蛋白的表达,而空白对照组和空白质粒转染组均未检测到hNIS mRNA和蛋白的表达。结论脂质体法可有效地将hNIS基因转染至SW480细胞并成功表达hNIS蛋白。Objective To transfect human sodium/iodide symporter gene hNIS into colon cancer SW480 cells and to ex- amine the hNIS mRNA and protein expression. Methods The recombinant plasmid （pcDNA3.1^＋-hNIS） was constructed, identi- fied and transfected into SW480 cells with lipofectamine 2000. Reverse transcription-polymerase chain reaction （RT-PCR） and Western blot were applied to detect the hNIS mRNA and protein expression. Results Enzyme digestion and DNA sequencing confirmed the size and direction of the recombinant （pcDNA3.1＋-hNIS） plasmid. RT-PCR and Western blot showed the expres- sion of hNIS mRNA and protein in pcDNA3.1＋-hNIS-transfected SW480 cells, not in non-transfected cells or blank plas- mid-transfected cells. Conclusion The hNIS gene is effectively transfected into human colon cancer SW480 cells and the NIS protein is expressed successfully.

关 键 词：钠 碘转运体 蛋白免疫印迹 基因转染 脂质体 

分 类 号：R735.3[医药卫生—肿瘤]