Hath1基因的重组腺病毒转染新生小鼠耳蜗成纤维细胞的初步研究  

Human hath1 gene recombinant adenovirus transfected mouse cochlea fibroblast

作　　者：陈杰[1] 高下[1] 

机构地区：[1]南京大学医学院附属南京鼓楼医院耳鼻咽喉科,江苏南京210008

出　　处：《中国现代医学杂志》2014年第6期24-27,共4页China Journal of Modern Medicine

基　　金：国家自然科学基金面上项目(No:30973302);江苏省科教兴卫工程医学重点人才基金(No:RC2007010);南京市医学重点科技发展项目(No:201108019)

摘　　要：目的检测含有人Hath1基因的重组腺病毒Ad-DsRed2-Hath1对新生小鼠耳蜗成纤维细胞的转染情况及Hath1基因在新生小鼠耳蜗成纤维细胞中的表达情况。方法培养新生小鼠耳蜗成纤维细胞,用含有目的基因Hath1的重组腺病毒Ad-DsRed2-Hath1对其进行转染,转染后24 h通过荧光显微镜观察新生小鼠耳蜗成纤维细胞的转染情况,并通过免疫荧光的方法检测Hath1基因在新生小鼠耳蜗成纤维细胞中的表达情况。结果含有人Hath1基因的重组腺病毒Ad-DsRed2-Hath1能高效转染新生小鼠耳蜗成纤维细胞,Hath1基因在新生小鼠耳蜗成纤维细胞中能有效表达。结论含有人Hath1基因的重组腺病毒Ad-DsRed2-Hath1对新生小鼠耳蜗成纤维细胞的高效转染和有效表达为研究Hath1基因转染对正常及致聋小鼠耳蜗效应的实验奠定了基础。[ Objective ] To detect the transfection and expression of Ad-DsRed2-Hathl in mouse cochlea fibrob- last. [Methods] After Ad-DsRed2-Hathl had been abstracted successfully, it was transfected into mouse cochlea fibroblast. Twenty four hours later, the efficiency of the transfection was analyzed by fluorescence microscope and the expression of Hathl gene in mouse cochlea fibroblast was detected by the means of immunofluorescence. [Results] The Ad-DsRed2-Hathl could transfect into mouse cochlea fibroblast effectively, and Hathl could express properly. The transfected fibroblasts displaying green fluorescence were observed under fluorescence microscope. The im- munofluorescence results showed that positive reactant of green fluorescence could be seen in the cytoplasm of fi- broblasts transfected by Ad-DsRed2-Hathl, while the no Hathl expression was observed in the fibroblasts trans- fected by Ad-DsRed2. [ Conclusions ] The effective transfection of Ad-DsRed2-Hathl into mouse cochlea fibroblast and proper expression of Hathl in transfected cells laid the basis for the following experiments, such as Hathl gene transfection in deaf or normal cochlea and so on.

关 键 词：Hath1 腺病毒 基因转染 耳蜗 成纤维细胞 小鼠 

分 类 号：R764.35[医药卫生—耳鼻咽喉科]