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The role of endoplasmic reticular Ca2+ stores in cell viability and tumor necrosis factor-α production of the murine macrophage RAW 264.7 cell line

  • Original Paper
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Journal of Biomedical Science

Abstract

Thapsigargin (TG), an endoplasmic reticular (ER) Ca2+-ATPase inhibitor, can increase the intracellular calcium concentration and then deplete the TG-sensitive intracellular Ca2+ pool. In this study, we investigated the effects of TG on cell viability and tumor necrosis factor-α (TNF-α) production in the murine macrophage RAW 264.7 cell line. We found that treatment with TG (10–800 nM) induced apoptosis in RAW 264.7 cells in a dose-dependent manner (IC50, 200 nM). Lipopolysaccharide (LPS, 1 µg/ml) markedly potentiated low concentrations of TG (10–75 nM) in inducing apoptosis (IC50, 20 nM) as revealed by the DNA ladder. Polymycin B (an LPS receptor antagonist) inhibited the cytotoxic effect induced by LPS plus TG. Although TG, A23187 and ionomycin all definitely increased intracellular Ca2+ concentrations, neither A23187 nor ionomycin mimicked TG in inducing apoptotic events in LPS-activated RAW 264.7 cells. Moreover, the production of TNF-α induced by LPS was profoundly potentiated by TG but not by A23187 or by ionomycin. We conclude from these combined results that TG-sensitive ER Ca2+ stores play a pivotal role in modulating cell viability and TNF-α production. The mutual potentiation between the LPS receptor signaling pathway and the depletion of ER Ca2+ stores implies the existence of cross-talk between these multiregulatory mechanisms in this murine macrophage RAW 264.7 cell line.

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Authors and Affiliations

  1. Institute of Pharmacology, College of Medicine, National Taiwan University, Taipei, Taiwan

    Shoei-Yn Lin-Shiau

  2. Institute of Toxicology, College of Medicine, National Taiwan University, No. 1, Section 1, Jen-Ai Road, 100, Tapei, Taiwan

    Yu-Jen Chen &  Shoei-Yn Lin-Shiau

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  1. Yu-Jen Chen
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Chen, YJ., Lin-Shiau, SY. The role of endoplasmic reticular Ca2+ stores in cell viability and tumor necrosis factor-α production of the murine macrophage RAW 264.7 cell line. J Biomed Sci 7, 122–127 (2000). https://doi.org/10.1007/BF02256618

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