Development and evaluation of HEK293 cells stably expressing CRFR1
10.3969/j.issn.1001-1978.2014.08.015
- VernacularTitle:稳定表达CRFR1的HEK293细胞株建立与功能鉴定
- Author:
Zhijuan GE
;
Jinmei YU
;
Xiaoyun MA
;
Xiaoyan LIU
;
Jianquan ZHENG
- Publication Type:Journal Article
- Keywords:
CRF;
CRFR1;
HEK293 cells;
transfec-tion;
cAMP;
GPCR
- From:
Chinese Pharmacological Bulletin
2014;(8):1113-1115,1116
- CountryChina
- Language:Chinese
-
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.
