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J. Biol. Chem., Vol. 280, Issue 25, 24221-24226, June 24, 2005

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Nuclear Phospholipase C 1 (PLC1) Affects CD24 Expression in Murine Erythroleukemia Cells^*

Roberta Fiume, Irene Faenza, Alessandro Matteucci, Annalisa Astolfi¶, Marco Vitale||, Alberto Maria Martelli**, and Lucio Cocco

From the Department of Anatomical Sciences, Cellular Signaling Laboratory, University of Bologna, 40126 Bologna, Italy, the ^**School of Pharmacy, University of Bologna, 40126 Bologna, Italy, ITOI-CNR, Unit of Bologna, c/o IOR 40136 Bologna, Italy, the ^¶Department of Experimental Pathology, Cancer Research Section, University of Bologna, 40126 Bologna, Italy, and the ^||Department of Anatomy, Pharmacology and Forensic Medicine, Human Anatomy Section, University of Parma, 43100 Parma, Italy

Inositide-specific phospholipase C (PLC) 1 is a key enzyme in nuclear lipid signal transduction affecting cell cycle progression and may be directly involved in regulation of gene expression and hematopoiesis. By microarrays, we compared the effect of nuclear PLC1 overexpression with that of PLC M2b cytoplasmatic mutant, which is exclusively located in the cytoplasm, in murine erythroleukemia cells. Out of 9000 genes analyzed, the CD24 gene, coding for an antigen involved in differentiation and hematopoiesis as well, was up-regulated in cells overexpressing nuclear PLC1 as compared with both cells overexpressing the M2b cytoplasmatic mutant and the wild type cells. Here we show that nuclear PLC1 up-regulated the expression of CD24. The correlation was strengthened by the observation that when PLC1 expression was silenced by means of small interfering RNA, CD24 expression was down-regulated. We also demonstrated that PLC1-dependent up-modulation of CD24 was mediated, at least in part, at the transcriptional level, in that PLC1 affected the CD24 promoter activity. Moreover, the up-regulation of CD24 was higher during erythroid differentiation of murine erythroleukemia cells. Altogether our findings, obtained by combining microarrays, phenotypic analysis, and small interfering RNA technology, identify CD24 as an molecular effector of nuclear PLC1 signaling pathway in murine erythroleukemia cells and strengthen the contention that nuclear PLC1 constitutes a key step in erythroid differentiation in vitro.

Received for publication, October 18, 2004 , and in revised form, April 7, 2005.

^* This work was supported by Italian FIRB and Cofin from MIUR, Italian Association for Cancer Research, CNR-MIUR Oncology Project and CARISBO Foundation. 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.

To whom correspondence should be addressed: Dept. of Anatomical Sciences, University of Bologna, Via Irnerio 48, I-40126 Bologna, Italy. Tel.: 39-051-244467; Fax: 39-051-251735; E-mail: lcocco{at}biocfarm.unibo.it.

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