Introduction and objectives Pancreatic adenocarcinoma (PDAC) is initiated and sustained by a subpopulation of cells defined as cancer stem cells (PCSCs). PCSCs are responsible for pancreatic tumorigenesis, tumour progression, and resistance to chemotherapeutic treatment.The aim of this work is the identification of new biomarkers or diagnostic/therapeutic targets of PDAC, through a proteomic comparison of a PDAC cell line and the derived PCSCs. Methods In this study, we performed an iTRAQ 8-plex/2D-LC-MS/MS analysis to compare the secretomes and whole proteomes of Panc1 and PCSCs cell lines. Results and Discussion The iTRAQ approach allowed us to identify 1146 proteins. Out of these, 111 proteins were found have higher abundance in the conditioned media as compared to the whole cell lysate of Panc1 and PCSCs cells. By using secretion prediction tools, we have found that 25% of the 111 proteins is exported by the classical secretory pathways, while the 75% is secreted by non-classical pathways. Then we performed a quantitative comparison of Panc1 and PCSCs secretomes, and whole proteome, identifying 71 and 162 differentially expressed proteins respectively. The global implications of secreted and intracellular differentially regulated proteins were elucidated by the Ingenuity Pathway Analysis (IPA). The differentially expressed proteins are involved in cell proliferation, invasion and contribute to epithelial-mesenchymal transition (EMT). Conclusions Our results indicate that iTRAQ technology is a promising way to investigate the PCSC biological model and to find useful biomarkers for PDAC.

SHOTGUN PROTEOMIC ANALYSIS OF PANCREATIC CANCER STEM CELLS

BRANDI, JESSICA;DALLA POZZA, Elisa;DANDO, Ilaria;Biondani, Giulia;PALMIERI, Marta;CECCONI, Daniela
2014

Abstract

Introduction and objectives Pancreatic adenocarcinoma (PDAC) is initiated and sustained by a subpopulation of cells defined as cancer stem cells (PCSCs). PCSCs are responsible for pancreatic tumorigenesis, tumour progression, and resistance to chemotherapeutic treatment.The aim of this work is the identification of new biomarkers or diagnostic/therapeutic targets of PDAC, through a proteomic comparison of a PDAC cell line and the derived PCSCs. Methods In this study, we performed an iTRAQ 8-plex/2D-LC-MS/MS analysis to compare the secretomes and whole proteomes of Panc1 and PCSCs cell lines. Results and Discussion The iTRAQ approach allowed us to identify 1146 proteins. Out of these, 111 proteins were found have higher abundance in the conditioned media as compared to the whole cell lysate of Panc1 and PCSCs cells. By using secretion prediction tools, we have found that 25% of the 111 proteins is exported by the classical secretory pathways, while the 75% is secreted by non-classical pathways. Then we performed a quantitative comparison of Panc1 and PCSCs secretomes, and whole proteome, identifying 71 and 162 differentially expressed proteins respectively. The global implications of secreted and intracellular differentially regulated proteins were elucidated by the Ingenuity Pathway Analysis (IPA). The differentially expressed proteins are involved in cell proliferation, invasion and contribute to epithelial-mesenchymal transition (EMT). Conclusions Our results indicate that iTRAQ technology is a promising way to investigate the PCSC biological model and to find useful biomarkers for PDAC.
Shotgun Proteomics
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11562/830365
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