Survivin基因siFNA表达载体的构建及其在食管癌EC109细胞中的表达

目的为研究survivin基因的功能及食管癌的基因治疗,构建针对survivin基因的siRNA的表达载体,转染细胞EC109后观察其对survivm基因表达的抑制作用.方法利用网站选择适当位点设计并合成针对survivin基因nRNA的寡核苷酸序列,插入质粒pBAsi-hU6中形成重组载体pBAsi-survlvin.重组载体经测序鉴定后转染食管癌细胞EC109,western blot检测转染前后survivin蛋白的表达情况.结果测序结果与所合成的寡核苷酸完全一致,证实成功构建了针对sLurvivin基因的siRNA表达载体.Westem blot检测显示转染食管癌细胞EC109后有效抑制了survivin基因的表达.结论针对surivivin基因的siRNA表达载体成功构建,并能有效抑制食管癌细胞EC109中survivin基因的表达.

Construction of survivin siRNA expression vector and its expression in esophagus carcinoma cell line EC109

Objective] The aim of this study was to construct a siRNA expression plasmid against gene survivin.So we can assess its function on esophagus carcinoma cell EC109.Methods] Potential RNAi oligonucleotides of survivin were selected on appropriate site.Then a survivin siRNA plasmid was constructed using a pBAsi-hU6 vector.The plasmid was sequenced to confirm the inserted sequence.Western blot analysis was taken to know about the change of survivin protein after the constructed plasmid had been transfected into EC109 cells.Results] It was confirmed by sequencing that the plasmid had been constructed successfully.The result of Western blot showed that survivin siRNA plasmid had inhibited survivin expression in EC109 cells obviously.Conclusion] The survivin siRNA plasmid was constructed successfully.Survivin protein expression of EC109 was inhibited by RNAi.