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J. Biol. Chem., Vol. 280, Issue 27, 25830-25839, July 8, 2005

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Search of Factors That Intermediate Cytokine-induced Group IIA Phospholipase A₂ Expression through the Cytosolic Phospholipase A₂- and 12/15-Lipoxygenase-dependent Pathway^*

Hiroshi Kuwata, Takuya Nonaka, Makoto Murakami, and Ichiro Kudo

From the Department of Health Chemistry, School of Pharmaceutical Sciences, Showa University, 1-5-8 Hatanodai, Shinagawa-ku, Tokyo 142-8555, Japan

Inducible expression of group IIA secretory phospholipase A₂ (sPLA₂-IIA) by interleukin-1 (IL-1) and tumor necrosis factor- (TNF) is under the control of group IVA cytosolic PLA₂ and 12/15-lipoxygenase (12/15-LOX) in rat fibroblastic 3Y1 cells. We show here that this cytokine induction of sPLA₂-IIA mRNA requires de novo protein synthesis. By means of cDNA array analysis, we found that the level of the CXC chemokine MIP-2 (macrophage inflammatory protein-2) was significantly elevated in 12/15-LOX-transfected cells compared with control cells. IL-1/TNF-stimulated induction of endogenous MIP-2 preceded that of sPLA₂-IIA, and exogenous MIP-2 induced sPLA₂-IIA dose-dependently. Moreover, a MIP-2-specific antisense oligonucleotide and small interfering RNA attenuated the IL-1/TNF-induced expression of sPLA₂-IIA, suggesting that MIP-2 is an absolute intermediate requirement for optimal induction of sPLA₂-IIA. In addition, the expression of c-jun and fra-1, which are components of the transcription factor AP-1, was elevated in 12/15-LOX-transfected cells, in which cytokine-dependent binding of AP-1 to the sPLA₂-IIA promoter was increased significantly. Conversely, the receptors for transforming growth factor- and platelet-derived growth factor, which contributed to down-regulation of sPLA₂-IIA expression, were decreased following 12/15-LOX overexpression. Taken together, 12/15-LOX-dependent up-regulation of sPLA₂-IIA expression may result from the interplay between accelerated MIP-2 signaling, AP-1 activation, and attenuated transforming growth factor- and platelet-derived growth factor signaling.

Received for publication, January 5, 2005 , and in revised form, April 8, 2005.

^* This work was supported by grants-in-aid for scientific research from the Ministry of Education, Science, Culture, Sports, and Technology of Japan. 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.

Present address: The Tokyo Metropolitan Institute of Medical Science, 3-18-22 Honkomagome, Bunkyo-ku, Tokyo 113-8613, Japan.

To whom correspondence should be addressed: Dept. of Health Chemistry, School of Pharmaceutical Sciences, Showa University, 1-5-8 Hatanodai, Shinagawa-ku, Tokyo 142-8555, Japan. Tel.: 81-3-3784-8196; Fax: 81-3-3784-8245; E-mail: kudo{at}pharm.showa-u.ac.jp.

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