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 Blood, 1 April 2007, Vol. 109, No. 7, pp. 3069-3075.
Prepublished online as a Blood First Edition Paper on December 19, 2006; DOI 10.1182/blood-2006-08-043257.

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NEOPLASIA

Overexpression of nucleolin in chronic lymphocytic leukemia cells induces stabilization of bcl2 mRNA

Yoko Otake1, Sridharan Soundararajan1, Tapas K. Sengupta1, Ebenezer A. Kio2, James C. Smith2, Mauricio Pineda-Roman2, Robert K. Stuart2, Eleanor K. Spicer1, and Daniel J. Fernandes1

1 The Department of Biochemistry and Molecular Biology and the 2 Division of Hematology/Oncology of the Department of Medicine, Medical University of South Carolina, Charleston, SC

B-cell chronic lymphocytic leukemia (CLL) is characterized by the accumulation of clonal B cells that are resistant to apoptosis as a result of bcl2 oncogene overexpression. Studies were done to determine the mechanism for the up-regulation of bcl-2 protein observed in CD19+ CLL cells compared with CD19+ B cells from healthy volunteers. The 11-fold higher level of bcl-2 protein in CLL cells was positively correlated with a 26-fold elevation in the cytosolic level of nucleolin, a bcl2 mRNA–stabilizing protein. Measurements of the bcl2 heterogeneous nuclear/bcl2 mRNA (hnRNA)/mRNA ratios and the rates of bcl2 mRNA decay in cell extracts indicated that the 3-fold higher steady-state level of bcl2 mRNA in CLL cells was the result of increased bcl2 mRNA stability. Nucleolin was present throughout the nucleus and cytoplasm of CLL cells, whereas in normal B cells nucleolin was only detected in the nucleus. The addition of recombinant human nucleolin to extracts of normal B cells markedly slowed the rate of bcl2 mRNA decay. SiRNA knockdown of nucleolin in MCF-7 cells resulted in decreased levels of bcl2 mRNA and protein but no change in ß-actin. These results indicate that bcl-2 overexpression in CLL cells is related to stabilization of bcl2 mRNA by nucleolin.


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