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Dexamethasone induces the secretion of annexin I in immature lymphoblastic cells by a calcium-dependent mechanism

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Abstract

The mechanisms by which glucocorticoids (GC) regulate annexin I (ANXA1) secretion in different cells are still a matter of debate. The aims of this study were to evaluate the ability of dexamethasone (Dex) to induce ANXA1 secretion and to investigate the roles of the intracellular free Ca2+ concentration ([Ca2+]i), and of the GC receptor, on that process. For this purpose, the human immature lymphoblastic CCRF-CEM cell line was used. Treatment of the cells with Dex, for up to 4 h, significantly reduced the intracellular content of ANXA1 and increased the amount of this protein bound to the outer surface of the plasma membrane, whereas exposure of cells to Dex, for 12 h, induced the synthesis of ANXA1. At the same short time periods, Dex also induced a significant increase in the [Ca2+]i. Incubation of the cells with BAPTA-AM (10 μM), a cell-permeant high affinity Ca2+ chelator, completely inhibited Dex-induced ANXA1 secretion. Furthermore, the Ca2+ ionophore, ionomycin, alone induced ANXA1 cleavage, but not its secretion. Additionally, we used brefeldin A to investigate the involvement of the classical endoplasmic reticulum (ER)-Golgi pathway of protein secretion in the release of ANXA1. The GC receptor antagonist, RU486, neither reverted the Dex-dependent ANXA1 secretion nor inhibited the increase of the [Ca2+]i induced by Dex. Together, our results indicate that Dex induces ANXA1 synthesis and secretion in CCRF-CEM cells. ANXA1 secretion in this cell type show the following characteristics: (i) is unlikely to involve the classical ER-Golgi pathway; (ii) requires a Ca2+-dependent cleavage of ANXA1; (iii) involves both Ca2+-dependent and independent mechanisms; and (iv) is apparently independent of the GC receptor alpha isoform.

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Authors and Affiliations

  1. Center for Neuroscience of Coimbra, Department of Zoology, University of Coimbra, 3004-517, Coimbra, Portugal

    M. Castro-Caldas, C.B. Duarte & A.P. Carvalho

  2. Center for Neuroscience of Coimbra, Department of Zoology and Faculty of Pharmacy, University of Coimbra, 3004-517, Coimbra, Portugal

    M.C. Fernandes Lopes

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  1. M. Castro-Caldas
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  2. C.B. Duarte
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  3. A.P. Carvalho
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  4. M.C. Fernandes Lopes
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Castro-Caldas, M., Duarte, C., Carvalho, A. et al. Dexamethasone induces the secretion of annexin I in immature lymphoblastic cells by a calcium-dependent mechanism. Mol Cell Biochem 237, 31–38 (2002). https://doi.org/10.1023/A:1016502120139

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