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J. Biol. Chem., Vol. 280, Issue 30, 28152-28161, July 29, 2005

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CTCF Regulates Growth and Erythroid Differentiation of Human Myeloid Leukemia Cells^*

Verónica Torrano, Igor Chernukhin¶, France Docquier¶, Vivien D'Arcy¶, Javier León, Elena Klenova¶||**, and M. Dolores Delgado||

From the Grupo de Biología Molecular del Cáncer, Departamento de Biologia Molecular, Unidad de Biomedicina-CSIC, Universidad de Cantabria, 39011 Santander, Spain and the ^¶Department of Biological Sciences, Central Campus, University of Essex, Wivenhoe Park, Colchester, Essex CO4 3SQ, United Kingdom

CTCF is a transcription factor and a candidate tumor suppressor that contains a DNA-binding domain composed of 11 zinc fingers. We reported previously that CTCF is differentially regulated during differentiation of human myeloid leukemia cells. In this study we aimed to investigate the role of CTCF in myeloid cell differentiation. A human cell line, K562, that can be chemically induced to differentiate into various hematopoietic lineages was chosen as a model system for this study. Several K562 cell lines with constitutive and conditional expression of CTCF have been generated. By using these model systems we demonstrated that: (i) ectopic expression of CTCF in K562 cells led to growth retardation and promotion of differentiation into the erythroid lineage; (ii) CTCF knock-down significantly inhibited differentiation of K562 cells into erythroid lineage; (iii) differentiation of K562 into the megakaryocytic lineage was not significantly affected; and (iv) down-regulation of MYC has been identified as one of the mechanisms by which CTCF promotes erythroid differentiation. Taken together our results demonstrate that CTCF is involved in the control of myeloid cell growth and differentiation.

Received for publication, February 8, 2005 , and in revised form, May 31, 2005.

^* This work was supported in part by Grants FIS01-1129 and FISPI041083 from the Spanish Ministerio de Sanidad y Consumo (to M. D. D.), Grant SAF02-4193 from Spanish Ministerio de Educación (to J. L.), a Biomedical Research Collaboration Grant from the Wellcome Trust (to E. K., M. D. D., and J. L.), the Breast Cancer Campaign (to F. D. and E. K.), and the Research Promotion Fund from the University of Essex (to I. C., F. D., and E. K.). 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.

Supported by a predoctoral fellowship from the Fondo Investigacion Sanitaria (FIS) and the University of Cantabria.

^|| Both authors contributed equally to this article.

^** To whom correspondence may be addressed: Dept. Biological Sciences, University of Essex, Wivenhoe Park, Colchester, Essex CO4 3SQ, United Kingdom. Tel.: 44-1206-874868; Fax: 44-1206-872592; E-mail: klenovae{at}essex.ac.uk.

To whom correspondence may be addressed: Dept. Biología Molecular, Facultad de Medicina, 39011-Santander, Spain. Tel.: 34-942-201955; Fax: 34-942-201945; E-mail: delgadmd{at}unican.es.

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