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Originally published In Press as doi:10.1074/jbc.M413076200 on January 4, 2005

J. Biol. Chem., Vol. 280, Issue 10, 8686-8693, March 11, 2005
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Expression and Function of the Nuclear Factor of Activated T Cells in Colon Carcinoma Cells

INVOLVEMENT IN THE REGULATION OF CYCLOOXYGENASE-2*

Javier Duque, Manuel Fresno, and Miguel A. Iñiguez{ddagger}

From the Centro de Biología Molecular "Severo Ochoa," Departamento de Biología Molecular, Universidad Autónoma de Madrid, 28049 Madrid, Spain

Increasing evidence shows a crucial role of the Ca2+/ calcineurin-mediated activation of the nuclear factor of activated T cells (NFAT) in the regulation of a variety of processes in nonimmune cells. Here we provide evidence that NFATc1 and NFATc2 are expressed in human colon carcinoma cell lines. These proteins are translocated from the cytoplasm to the nucleus upon treatment with a combination of phorbol 12-myristate 13-acetate plus the calcium ionophore A23187. Subsequent to translocation to the nucleus, NFATc1 and NFATc2 were able to bind to a NFAT response element in the DNA, regulating transcriptional activation of genes containing a NFAT-responsive element such as cyclooxygenase-2 (COX-2). COX-2 expression and prostaglandin E2 (PGE2) production were induced upon pharmacological stimuli leading to NFAT activation and blunted by inhibition of calcineurin phosphatase with cyclosporin A or tacrolimus (FK506). Expression of NFAT wild type protein or the active catalytic subunit of calcineurin transactivates COX-2 promoter activity, whereas a dominant negative mutant of NFAT inhibited COX-2 induction in colon carcinoma cell lines. Furthermore, mutation or deletion of NFAT binding sites in the human COX-2 promoter greatly diminished its induction by phorbol 12-myristate 13-acetate/calcium ionophore A23187. These findings demonstrate the presence and activation of NFAT in human colon carcinoma cells, with important implications in the regulation of genes involved in the transformed phenotype as COX-2.

Received for publication, November 19, 2004 , and in revised form, December 27, 2004.

* This work was supported by Ministerio de Ciencia y Tecnología Grant BMC2001-0177, Ministerio de Educación y Ciencia-FEDER Grant BFU2004-04157, RECAVA cardiovascular network (C03/01) from Fondo de Investigaciones Sanitarias, EICOSANOX integrated project and MAIN network of excellence from the 6th European Union Framework Programme, Laboratorios del Dr. ESTEVE, and Comunidad de Madrid (08.3/0007/1). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

{ddagger} Recipient of the Ramón y Cajal Program of the Ministerio de Ciencia y Tecnología of Spain. To whom correspondence should be addressed. Tel.: 34-914978410; Fax: 34-914974799; E-mail: Mainiguez{at}cbm.uam.es.


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