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Identification of phosphoproteins as possible differentiation markers in all-trans-retinoic acid-treated neuroblastoma cells

Mandili, Giorgia and Marini, Cristina and Carta, Franco and Zanini, Cristina and Prato, Mauro and Khadjavi, Amina and Turrini, Francesco Michelangelo and Giribaldi, Giuliana (2011) Identification of phosphoproteins as possible differentiation markers in all-trans-retinoic acid-treated neuroblastoma cells. PLoS One, Vol. 6 (5), e18254. ISSN 1932-6203. Article.

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DOI: 10.1371/journal.pone.0018254

Abstract

Background: Neuroblastic tumors account for 9-10% of pediatric tumors and neuroblastoma (NB) is the first cause of death in pre-school age children. NB is classified in four stages, depending on the extent of spreading. A fifth type of NB, so-called stage 4S (S for special), includes patients with metastatic tumors but with an overall survival that approximates 75% at five years. In most of these cases, the tumor regresses spontaneously and regression is probably associated with delayed neuroblast cell differentiation.
Methodology/Principal Findings: In order to identify new early markers to follow and predict this process for diagnostic and therapeutics intents, we mimicked the differentiation process treating NB cell line SJ-NK-P with all-trans-retinoic acid (ATRA) at different times; therefore the cell proteomic pattern by mass spectrometry and the phosphoproteomic pattern by a 2-DE approach coupled with anti-phosphoserine and anti-phosphotyrosine western blotting were studied.
Conclusions/Significance: Proteomic analysis identified only two proteins whose expression was significantly different in treated cells versus control cells: nucleoside diphosphate kinase A (NDKA) and reticulocalbin-1 (RCN1), which were both downregulated after 9 days of ATRA treatment. However, phosphoproteomic analysis identified 8 proteins that were differentially serine-phosphorylated and 3 that were differentially tyrosine-phosphorylated after ATRA treatment. All proteins were significantly regulated (at least 0.5-fold down-regulated). Our results suggest that differentially phosphorylated proteins could be considered as more promising markers of differentiation for NB than differentially expressed proteins.

Item Type:Article
ID Code:6238
Status:Published
Refereed:Yes
Uncontrolled Keywords:Endoplasmic-reticulum stress, human hepatocarcinoma cells, glucose-regulated protein, neuronal differentiation, cancer-cells, neurite outgrowth, prognostic marker, Ewings-sarcoma, SH-SY5Y cells, Kinase-C
Subjects:Area 05 - Scienze biologiche > BIO/10 Biochimica
Divisions:002 Altri enti e centri di ricerca del Nord Sardegna > Nurex Bioresearch Sassari, Sassari
Publisher:Public Library of Science
ISSN:1932-6203
Copyright Holders:© 2011 Mandili et al.
Deposited On:11 Jul 2011 10:30

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