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- Title
The functional expression of calcium-sensing receptor in the differentiated THP-1 cells.
- Authors
Xi, Yu-hui; Li, Hong-zhu; Zhang, Wei-hua; Wang, Li-na; Zhang, Li; Lin, Yan; Bai, Shu-zhi; Li, Hong-xia; Wu, Ling-yun; Wang, Rui; Xu, Chang-qing
- Abstract
The expression and function of calcium-sensing receptor (CaSR) in differentiated THP-1 (human acute monocytic leukemia cell line) cells are unknown currently. This study investigated above-mentioned issues using TRAP staining, immunofluorescence staining, Western blotting, ELISA, and Laser Confocal Scanning Microscopy techniques. We found that CaSR protein was expressed, and mainly located in the membrane and cytoplasm in differentiated THP-1 cells. Elevated extracellular calcium or GdCl (an agonist of CaSR) raised intracellular calcium concentration. And this increase was inhibited or abolished by NPS2390 (an inhibitor of CaSR), U73122 (a specific inhibitor of phospholipase C, PLC) or thapsigargin (a Ca-ATPase inhibitor). The extracellular GdCl elevation stimulated both of IL-1β and TNFα release, and this effect of GdCl was inhibited by NPS2390. In conclusion, CaSR is functionally expressed in differentiated THP-1 cells, and the activated CaSR contributes to intracellular calcium increment through Gq-PLC- inositol triphosphate (IP3) pathway and commits to cytokine secretion. These results suggest that CaSR might be involved in a variety of pathological processes mediated by activated monocyte-macrophages.
- Subjects
GENE expression; INTRACELLULAR calcium; CELL differentiation; MONOCYTIC leukemia; MACROPHAGES; ENZYME-linked immunosorbent assay; ENZYME inhibitors; WESTERN immunoblotting; IMMUNOFLUORESCENCE
- Publication
Molecular & Cellular Biochemistry, 2010, Vol 342, Issue 1/2, p233
- ISSN
0300-8177
- Publication type
Article
- DOI
10.1007/s11010-010-0489-3