稳定表达miR-17-5p的PC12细胞株构建及鉴定  

Establishment and appraisal of miR-17-5p stably transfected PC12 cell line

作　　者：张皓云[1] 王鲁娟[2] 王凤斌[3] 刘红兵[3] 苏文霞[3] 

机构地区：[1]潍坊医学院基础医学概论教研室,山东潍坊261053 [2]潍坊医学院诊断学教研室,山东潍坊261053 [3]潍坊医学院生理学教研室,山东潍坊261053

出　　处：《实验技术与管理》2015年第12期66-68,共3页Experimental Technology and Management

基　　金：山东省自然科学基金资助项目(ZL2013HL065);山东省高等学校科技计划项目(J15LK10)

摘　　要：目的:构建稳定表达miR-17-5p的PC12细胞株。方法:将质粒PEGFP-N1-vector空载体和PEGFPN1-miR-17-5p表达载体扩增、抽提,利用脂质体转染技术将质粒转染PC12细胞,荧光显微镜观察绿色荧光蛋白GFP表达,Real-time PCR检测转染PC12细胞miR-17-5p的表达。结果:荧光显微镜观察到空载组和miR-17-5p组细胞都有绿色荧光蛋白高表达,Real-time PCR检测稳定转染miR 17-5p后PC12细胞内miR-17-5p的表达较空载组明显升高(P<0.01)。结论:miR-17-5p在PC12细胞稳定表达,为进一步研究miR-17-5p对神经细胞的调制作用提供有用的细胞模型。Objective：To establish stable expressing miR-17-5p PC12 cell line.Methods：Amplification and extraction of PEGFP-N1-vector and PEGFP-N1-miR-17-5p plasmids.Then the plasmids were transfected into PC12 cells by Lipofectamine 2000.Fluorescence microscope and real-time PCR were employed to detect the expression alterations of GFP and miR-17-5p in stable transfected PC12 cells.Results：High expressions of GFP were observed both in vector group and miR-17-5p group.Real-time PCR show that the expression of miR-17-5p is significantly elevated in PEGFP-N1-miR-17-5p stable transfected PC12 cells compared with the PEGFP-N1-vector stable transfected cells（P〈0.01）.Conclusion：Stable transfection of miR-17-5p PC12 cell line is established which will provide a useful tool to study the role of miR-17-5p in neurons.

关 键 词：微小RNA miR-17-5p PC12细胞 

分 类 号：R33-33[医药卫生—人体生理学] Q433[医药卫生—基础医学]