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Blood, 15 July 2008, Vol. 112, No. 2, pp. 394-397.
Prepublished online as a Blood First Edition Paper on April 23, 2008; DOI 10.1182/blood-2007-11-124065.
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Submitted November 14, 2007
Accepted March 17, 2008
SAGE analysis demonstrates increased expression of TOSO contributing to Fas mediated resistance in CLL
Rodrigo Proto-Siqueira, Rodrigo A Panepucci, Francisco P Careta, Abigail Lee, Andrew Clear, Kelly Morris, Carolyn Owen, Edgar G Rizzatti, Wilson A Silva-Jr, Roberto Passetto Falcao, Marco A Zago, and John G. Gribben*
Hematology Division and Center for Cell Based Therapy, Faculty of Medicine of Ribeirao Preto, University of Sao Paulo, Ribeirao Preto, Sao Paulo, Brazil
Institute of Cancer, Barts and The London School of Medicine and Dentistry, London, United Kingdom
* Corresponding author; email: john.gribben{at}cancer.org.uk.
To identify novel genes involved in the molecular pathogenesis of chronic lymphocytic leukemia (CLL) we performed a serial analysis of gene expression (SAGE) in CLL cells, and compared this with healthy B cells (nCD19+). We found a high level of similarity among CLL subtypes, but a comparison of CLL versus nCD19+ libraries revealed 55 genes that were over-represented and 49 genes that were down-regulated in CLL. A gene ontology analysis revealed that TOSO, which plays a functional role upstream of Fas extrinsic apoptosis pathway, was over-expressed in CLL cells. This finding was confirmed by real-time RT-PCR in 78 CLL and 12 nCD19+ cases (p<0.0001). We validated expression using flow cytometry and tissue microarray and demonstrated a 5.6 fold increase of TOSO protein in circulating CLL cells (p=0.013) and lymph nodes (p=0.0057). Our SAGE results have demonstrated that TOSO is a novel over-expressed anti-apoptotic gene in CLL.
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