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J. Biol. Chem., Vol. 280, Issue 20, 20086-20093, May 20, 2005

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CRD-BP/IMP1 Expression Characterizes Cord Blood CD34⁺ Stem Cells and Affects c-myc and IGF-II Expression in MCF-7 Cancer Cells^*

Panayotis Ioannidis, Louisa G. Mahaira, Sonia A. Perez¶, Angelos D. Gritzapis, Panagiota A. Sotiropoulou, Giannis J. Kavalakis||, Aris I. Antsaklis||, Constantin N. Baxevanis, and Michael Papamichail

From the Cancer Immunology Immunotherapy Center, Saint Savas Hospital, 171, Alexandras Avenue, Athens 115 22, Greece and ^||the 1st Obstetrics and Gynecology University Clinic, Alexandras Maternity Hospital, 80, Vassilissis Sofias Avenue, Athens 115 21, Greece

The coding region determinant-binding protein/insulin-like growth factor II mRNA-binding protein (CRD-BP/IMP1) is an RNA-binding protein specifically recognizing c-myc, leader 3' IGF-II and tau mRNAs, and the H19 RNA. CRD-BP/IMP1 is predominantly expressed in embryonal tissues but is de novo activated and/or overexpressed in various human neoplasias. To address the question of whether CRD-BP/IMP1 expression characterizes certain cell types displaying distinct proliferation and/or differentiation properties (i.e. stem cells), we isolated cell subpopulations from human bone marrow, mobilized peripheral blood, and cord blood, all sources known to contain stem cells, and monitored for its expression. CRD-BP/IMP1 was detected only in cord blood-derived CD34⁺ stem cells and not in any other cell type of either adult or cord blood origin. Adult BM CD34⁺ cells cultured in the presence of 5'-azacytidine expressed de novo CRD-BP/IMP1, suggesting that epigenetic modifications may be responsible for its silencing in adult non-expressing cells. Furthermore, by applying the short interfering RNA methodology in MCF-7 cells, we observed, subsequent to knocking down CRD-BP/IMP1, decreased c-myc expression, increased IGF-II mRNA levels, and reduced cell proliferation rates. These data 1) suggest a normal role for CRD-BP/IMP1 in pluripotent stem cells with high renewal capacity, like the CB CD34⁺ cells, 2) indicate that altered methylation may directly or indirectly affect its expression in adult cells, 3) imply that its de novo activation in cancer cells may affect the expression of c-Myc and insulin-like growth factor II, and 4) indicate that the inhibition of CRD-BP/IMP1 expression might affect cancer cell proliferation.

Received for publication, September 1, 2004 , and in revised form, March 11, 2005.

^* This work was supported by a grant from the Regional Operational Program Attika 20 (MIS code 59605GR (to M. P.) 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.

Contributed equally to this work.

^¶ To whom correspondence should be addressed: Cancer Immunology Immunotherapy Center, Saint Savas Hospital, 171 Alexandras Ave., 115 22 Athens, Greece. Tel.: 30-10-6409459; Fax: 30-10-6409516; E-mail: perez{at}ciic.gr.

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