Profiling B cell chronic lymphocytic leukemia by reverse phase protein array: Focus on apoptotic proteins.
B cell chronic lymphocytic leukemia (CLL) is characterized by the accumulation of B lymphocytes from proliferative activity and apoptosis resistance. The increased awareness of the importance of B cell receptor signaling in CLL has raised new opportunities for targeted intervention. Herein, we describe a study performed with the high-throughput RPPA (reverse phase protein array) technique, which allowed us to simultaneously study different molecules in a large series of patients. We analyzed B lymphocytes from 57 patients with CLL and 11 healthy subjects. Different pathways were assessed for activation/expression of key signaling proteins. Data obtained were validated by Western blotting and confocal microscopy. The RPPA investigation and its validation, identified 3 series of proteins: 1) molecules whose expression levels reached statistically significant differences in CLL vs. healthy controls (HSP70, Smac/DIABLO, cleaved PARP, and cleaved caspase-6); 2) proteins with a positive trend of difference in CLL vs. healthy controls (HS1, γ-tubulin, PKC α/β-II Thr-638/641, p38 MAPK Thr-180/Tyr-182, NF-κB Ser-536, Bcl2 Ser-70 and Src Tyr-527); and 3) molecules differentially expressed in patients with IGHV mutations vs. those without mutations (ZAP70, PKC-ζλ, Thr-410/403, and CD45). This study identified some molecules, particularly those involved in apoptosis control, which could be considered for further studies to design new therapeutic strategies in CLL.
Department of Medicine, Hematology and Clinical Immunology Branch, Padua University School of Medicine, Padova, Italy.|Venetian Institute of Molecular Medicine, Padova, Italy; and.
Federica
F
Frezzato
Department of Women's and Children's Health, University of Padua, Padova, Italy.
Benedetta
B
Accordi
Department of Medicine, Hematology and Clinical Immunology Branch, Padua University School of Medicine, Padova, Italy.|Venetian Institute of Molecular Medicine, Padova, Italy; and.
Valentina
V
Trimarco
Department of Medicine, Hematology and Clinical Immunology Branch, Padua University School of Medicine, Padova, Italy.|Venetian Institute of Molecular Medicine, Padova, Italy; and.
Cristina
C
Gattazzo
Department of Medicine, Hematology and Clinical Immunology Branch, Padua University School of Medicine, Padova, Italy.|Venetian Institute of Molecular Medicine, Padova, Italy; and.
Veronica
V
Martini
Department of Women's and Children's Health, University of Padua, Padova, Italy.
Gloria
G
Milani
Department of Women's and Children's Health, University of Padua, Padova, Italy.
Silvia
S
Bresolin
Department of Medicine, Hematology and Clinical Immunology Branch, Padua University School of Medicine, Padova, Italy.|Venetian Institute of Molecular Medicine, Padova, Italy; and.
Filippo
F
Severin
Department of Medicine, Hematology and Clinical Immunology Branch, Padua University School of Medicine, Padova, Italy.|Venetian Institute of Molecular Medicine, Padova, Italy; and.
Andrea
A
Visentin
Department of Women's and Children's Health, University of Padua, Padova, Italy.
Giuseppe
G
Basso
Department of Medicine, Hematology and Clinical Immunology Branch, Padua University School of Medicine, Padova, Italy; g.semenzato@unipd.it.|Venetian Institute of Molecular Medicine, Padova, Italy; and.
Gianpietro
G
Semenzato
Department of Medicine, Hematology and Clinical Immunology Branch, Padua University School of Medicine, Padova, Italy; livio.trentin@unipd.it.|Venetian Institute of Molecular Medicine, Padova, Italy; and.
Livio
L
Trentin
D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins / Q000032:analysis*; D001402:B-Lymphocytes / Q000737:chemistry* / Q000473:pathology; D015153:Blotting, Western; D020869:Gene Expression Profiling; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000032:analysis*; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D018613:Microscopy, Confocal; D009363:Neoplasm Proteins / Q000032:analysis*; D040081:Protein Array Analysis / Q000379:methods*
D051017:Apoptosis Regulatory Proteins; D047908:Intracellular Signaling Peptides and Proteins; D009363:Neoplasm Proteins
HSP70; Smac/DIABLO; lymphocyte; signal transduction