中国药理学通报Issue(8)：1113-1115,1116,4.DOI:10.3969/j.issn.1001-1978.2014.08.015
稳定表达CRFR1的HEK293细胞株建立与功能鉴定
Development and evaluation of HEK293 cells stably expressing CRFR1
摘要
Abstract
Aim ToconstructHEK293cellsstablyex-pressing corticotropin releasing factor receptor 1 ( CRFR1 ) , and evaluate its function by the cAMP as-say.Methods CulturedHEK293cellsweretransfect-ed with CRFR1-expressing vector by Lipofectamine 2000 and were selected by using G418 . CRFR1 ex-pression was detected by Western blot, RT-PCR and immunofluorescence.Results Westernblot,RT-PCR and immunofluorescence data revealed that the HEK293 cells expressed CRFR1 protein stably. The dose-responsive relationship experiment revealed that CRF induced a CRFR1-mediated cAMP production in HEK293 cells with EC50 =(5. 64 ± 0. 05) × 10 -10 mol ·L-1.Conclusion HEK293celllinesstablyex-pressing CRFR1 were constructed successfully, which would provide a cellular model to facilitate the research on the biological function of CRFR1 and CRFR1-targe-ted drug screening.关键词
促肾上腺皮质激素释放因子/CRFR1/HEK293 细胞/转染/cAMP/G蛋白偶联受体Key words
CRF/CRFR1/HEK293 cells/transfec-tion/cAMP/GPCR分类
医药卫生引用本文复制引用
葛治娟,于金梅,马晓芸,刘晓燕,郑建全..稳定表达CRFR1的HEK293细胞株建立与功能鉴定[J].中国药理学通报,2014,(8):1113-1115,1116,4.基金项目
国家科技部“重大新药创制”科技重大专项( No 2012ZX09301003-001) （ No 2012ZX09301003-001）
国家自然科学基金资助项目( No 31350001) （ No 31350001）
