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Volume 272, Number 9, Issue of February 28, 1997 pp. 5952-5958
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

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Activation of Cytosolic Phospholipase A₂ by Platelet-derived Growth Factor Is Essential for Cyclooxygenase-2-dependent Prostaglandin E₂ Synthesis in Mouse Osteoblasts Cultured with Interleukin-1

(Received for publication, October 2, 1996, and in revised form, December 2, 1996)

Qing-Rong Chen ^§ , Chisato Miyaura , Sayumi Higashi ^¶ , Makoto Murakami ^¶ , Ichiro Kudo ^¶ , Shigeru Saito ^§ , Takatoshi Hiraide ^§ , Yoshinobu Shibasaki ^§ and Tatsuo Suda

From the  Department of Biochemistry, School of Dentistry, ^¶ Department of Health Chemistry, School of Pharmaceutical Sciences, and ^§ Department of Orthodontics, School of Dentistry, Showa University, 1-5-8 Hatanodai, Shinagawa-ku, Tokyo 142, Japan

The synthesis of prostaglandins (PGs) is regulated by the arachidonic acid release by phospholipase A₂ (PLA₂) and its conversion to PGs by cyclooxygenase (COX). In the present study, we examined the regulation of PG synthesis by interleukin (IL)-1 in primary mouse osteoblastic cells isolated from mouse calvaria. Although IL-1 greatly enhanced cox-2 mRNA expression and its protein levels, PGE₂ was not produced until 24 h. When arachidonic acid was added to osteoblastic cells precultured with IL-1 for 24 h, PGE₂ was produced within 10 min. Of several growth factors tested, platelet-derived growth factor (PDGF) specifically initiated the rapid synthesis of PGE₂, which was markedly suppressed by a selective inhibitor of cox-2 (NS-398). In mouse osteoblastic cells, cytosolic PLA₂ (cPLA₂) mRNA and its protein were constitutively expressed and increased approximately 2-fold by IL-1, but secretory PLA₂ mRNA was not detected. PDGF rapidly stimulated PLA₂ activity, which was blocked completely by a cPLA₂ inhibitor (arachidonyltrifluoromethyl ketone). The PDGF-induced cPLA₂ activation was accompanied by phosphorylation of its protein. These results indicate that cox-2 induction by IL-1 is not sufficient, but cPLA₂ activation by PDGF is crucial for IL-1-induced PGE₂ synthesis in mouse osteoblasts.

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