四环素调控的介导bFGF重组腺相关病毒的构建及体外实验  

Construction of Doxycycline-inducible Adenoviral Vector-mediated Expression of Basic Fibroblast Growth Factor in Vitro

作　　者：宋珂[1] 陈美玲[1] 石琦[1] 青莹 曹颖光[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院口腔科,湖北武汉430030

出　　处：《口腔医学研究》2015年第9期851-855,共5页Journal of Oral Science Research

基　　金：国家自然科学基金(编号:81400490);湖北省自然科学基金(编号:2014CFB388)

摘　　要：目的:制备携带碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)的病毒载体,能被四环素调控表达。方法:构建重组质粒pAAV-S3-bFGF,生产重组病毒rAAV2-tet-off-bFGF,并感染小鼠颅骨前成骨细胞MC3T3-E1,实时定量PCR检测强力霉素(doxycycline,Dox)调控下rAAV2-tet-off-bFGF对MC3T3-E1中成骨相关因子的影响。结果:生产出高滴度及纯度的重组病毒rAAV2-tet-off-bFGF,能被Dox调控,感染MC3T3-E1后能促进胞内成骨相关因子的表达升高。结论:这种重组病毒rAAV2-tet-off-bFGF能在Dox调控下有效传递外源基因,为骨组织工程中的种子细胞提供基因传递载体。Objective： To establish the dox-inducible adenoviral vector and to evaluate its ability to deliver bFGF to ceils in bone tissue engineering. Methods： The adenoviral vector pAAV-S3-bFGF was constructed. Adenovirus was packaged in HEK293 cells and then infected the mouse osteoblastic cells （MC3T3-E1）. The efficiency of infection was evaluated by real-time PCR and Western blot. Results： The recombinant adenovirus rAAV2-tet- off--bFGF was regulated by Dox. MC3T3-E1 cells could be efficiently infected. Additionally, the expression of osteogenesis-related genes was significantly increased in vitro. Conclusion： The rAAV2-tet-off-bFGF can deliver exogenous genes efficiently into MC3T3-E1 cells, and has the potential to be used in bone tissue engineering.

关 键 词：腺相关病毒 基因传递 碱性成纤维细胞生长因子 四环素调控系统 

分 类 号：R96[医药卫生—药理学]