The functional expression of calcium-sensing receptor in the differentiated THP-1 cells.
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(3) (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(2+)-ATPase inhibitor). The extracellular GdCl(3) elevation stimulated both of IL-1beta and TNFalpha release, and this effect of GdCl(3) 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.
Xi
Yu-hui
YH
Department of Pathophysiology, Harbin Medical University, Harbin, China.
Li
Hong-zhu
HZ
Zhang
Wei-hua
WH
Wang
Li-na
LN
Zhang
Li
L
Lin
Yan
Y
Bai
Shu-zhi
SZ
Li
Hong-xia
HX
Wu
Ling-yun
LY
Wang
Rui
R
Xu
Chang-qing
CQ
D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism*; D002454:Cell Differentiation; D016207:Cytokines / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D043802:GTP-Binding Protein alpha Subunits, Gq-G11 / Q000378:metabolism; D005682:Gadolinium / Q000494:pharmacology; D020962:Heterotrimeric GTP-Binding Proteins / Q000378:metabolism; D006801:Humans; D007948:Leukemia, Monocytic, Acute / Q000378:metabolism* / Q000473:pathology*; D044169:Receptors, Calcium-Sensing / Q000378:metabolism*; D015398:Signal Transduction; D014407:Tumor Cells, Cultured; D010738:Type C Phospholipases / Q000378:metabolism
D000893:Anti-Inflammatory Agents; C095550:CASR protein, human; D016207:Cytokines; D044169:Receptors, Calcium-Sensing; D005682:Gadolinium; D010738:Type C Phospholipases; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D020962:Heterotrimeric GTP-Binding Proteins; C038958:gadolinium chloride; D002118:Calcium
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