Establishment and application of human CHO/NTR1 system.
- Author:
Guo ZHANG
;
Tao SUN
;
Hui-Juan LIU
;
Guo-Jun NIU
;
Feng XU
- Publication Type:Journal Article
- MeSH:
Animals;
CHO Cells;
Calcium Signaling;
Cricetinae;
Cricetulus;
Humans;
Pyrazoles;
pharmacology;
Quinolines;
pharmacology;
Receptors, Neurotensin;
antagonists & inhibitors;
genetics;
metabolism;
Transfection
- From:
Acta Pharmaceutica Sinica
2014;49(9):1273-1278
- CountryChina
- Language:Chinese
-
Abstract:
Neurotensin receptor-1 (NTR1), which can stimulate the intracellular cascade signal pathway, belongs to the large superfamily of G-protein coupled receptors. NTR1 is related to the occurrence and development of several kinds of diseases. In order to screen the inhibitors for the cancers associated with NTR1 protein, we established a CHO (Chinese hamster ovary) cell line in which human neurotensin receptor-1 was highly expressed. The method is to construct the recombinant plasmid which was lysed with the hNTR1 gene and transfect it into CHO cells. After selected with G418, the cell line was evaluated by Western blotting analysis and calcium flux assays. Through the calcium flux assays on FlexStation 3, we got the EC50 value of neurotensin peptide which is the natural NTR1 agonist, and the IC 50 value of SR48692 which is the known NTR1 antagonist. The established human CHO/NTR1 cell line can be used to study the profile of NTR1 biological activity and further screen of NTR1 antagonists and agonists.
