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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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Submitted August 24, 2006
Accepted November 8, 2006
Overexpression of nucleolin in chronic lymphocytic leukemia cells induces stabilization of bcl-2 mRNA
Yoko Otake, Sridharan Soundararajan, Tapas K Sengupta, Ebenezer A Kio, James C Smith, Mauricio Pineda-Roman, Robert K Stuart, Eleanor K Spicer, and Daniel J Fernandes*
Dept. of Biochemistry and Molecular Biology, and the Division of Hematology/Oncology, Dept of Medicine, Medical University of South Carolina, Charleston, SC
* Corresponding author; email: fernand{at}musc.edu.
B cell chronic lymphocytic leukemia (CLL) is characterized by the accumulation of clonal B cells that are resistant to apoptosis as a result of bcl-2 oncogene overexpression. Studies were done to determine the mechanism for the upregulation of bcl-2 protein observed in CD19+ CLL cells compared to CD19+ B cells from normal 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 bcl-2 mRNA stabilizing protein. Measurements of the bcl-2 hnRNA/mRNA ratios and the rates of bcl-2 mRNA decay in cell extracts indicated that the 3-fold higher steady-state level of bcl-2 mRNA in CLL cells was the result of increased bcl-2 mRNA stability. Nucleolin was present throughout the nucleus and cytoplasm of CLL cells, while 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 bcl-2 mRNA decay. SiRNA knockdown of nucleolin in MCF-7 cells resulted in decreased levels of bcl-2 mRNA and protein but no change in  -actin. These results indicate that bcl-2 overexpression in CLL cells is related to stabilization of bcl-2 mRNA by nucleolin.
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