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The functional expression of calcium-sensing receptor in the differentiated THP-1 cells

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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 GdCl3 (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 Ca2+-ATPase inhibitor). The extracellular GdCl3 elevation stimulated both of IL-1β and TNFα release, and this effect of GdCl3 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.

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Acknowledgments

This work was supported by the National Basic Research Program of China (973 Program, grant number 2007CB512000); National Natural Science Foundation of China (30871012, 30470688 and 30700288); Specialized Research Found for the Doctoral Program of Higher Education (20070226012); Harbin Medical University found for younger scientists (060016) and Technology Research Foundation of the Department of Education of Heilongjiang Province (11521078).

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Authors and Affiliations

  1. Department of Pathophysiology, Harbin Medical University, Harbin, 150086, China

    Yu-hui Xi, Hong-zhu Li, Wei-hua Zhang, Li-na Wang, Li Zhang, Yan Lin, Shu-zhi Bai, Hong-xia Li & Chang-qing Xu

  2. Bio-pharmaceutical Key Laboratory of Heilongjiang Province, Harbin, 150086, China

    Chang-qing Xu

  3. Department of Biology, Lakehead University, Thunder Bay, ON, P7B5E1, Canada

    Ling-yun Wu & Rui Wang

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  1. Yu-hui Xi
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Correspondence to Chang-qing Xu.

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Xi, Yh., Li, Hz., Zhang, Wh. et al. The functional expression of calcium-sensing receptor in the differentiated THP-1 cells. Mol Cell Biochem 342, 233–240 (2010). https://doi.org/10.1007/s11010-010-0489-3

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  • DOI: https://doi.org/10.1007/s11010-010-0489-3

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