Dataset Information

Human Serum LC-MS/MS - LC-MS/MS-based serum proteomics for identification of candidate biomarkers for hepatocellular carcinoma

ABSTRACT: Associating changes in protein levels with the onset of cancer has proven as an effective approach to identify clinically relevant diagnostic biomarkers. In the present study, we used sera from 205 patients representing two cohorts for biomarker discovery using label-free proteomic analysis by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). We performed first untargeted proteomic analysis of sera to identify candidate proteins with statistically significant difference between hepatocellular carcinoma (HCC) and patients with liver cirrhosis. These proteins were later on evaluated through targeted quantitation by multiple reaction monitoring (MRM) on a triple quadrupole mass spectrometer. The results of this study illustrate the power of combining untargeted and targeted quantitation methods for a comprehensive serum proteomic analysis, to investigate changes in protein levels and discover novel diagnostic biomarkers.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood Serum

DISEASE(S): Hepatocellular Carcinoma

SUBMITTER: Habtom Ressom

LAB HEAD: Habtom W. Ressom

PROVIDER: PXD001171 | Pride | 2015-06-02

REPOSITORIES: Pride

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			Action	DRS
	Egypt_002_020514_01.mzXML	Mzxml
	Egypt_003_020514_01.mzXML	Mzxml
	Egypt_004_020514_01.mzXML	Mzxml
	Egypt_005_020514_01.mzXML	Mzxml
	Egypt_007_020514_01.mzXML	Mzxml

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Publications

LC-MS/MS-based serum proteomics for identification of candidate biomarkers for hepatocellular carcinoma.

Tsai Tsung-Heng TH Song Ehwang E Zhu Rui R Di Poto Cristina C Wang Minkun M Luo Yue Y Varghese Rency S RS Tadesse Mahlet G MG Ziada Dina Hazem DH Desai Chirag S CS Shetty Kirti K Mechref Yehia Y Ressom Habtom W HW

Proteomics 20150429 13

Associating changes in protein levels with the onset of cancer has been widely investigated to identify clinically relevant diagnostic biomarkers. In the present study, we analyzed sera from 205 patients recruited in the United States and Egypt for biomarker discovery using label-free proteomic analysis by LC-MS/MS. We performed untargeted proteomic analysis of sera to identify candidate proteins with statistically significant differences between hepatocellular carcinoma (HCC) and patients with ...[more]

PMID: 25778709

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Project description:Background & Aims: Extracellular vesicles (EVs) play a pivotal role in connecting tumor cells and their local and distant microenvironment. Here, we aimed to understand the role and molecular basis of patient-derived EVs in modulating cancer stemness and tumorigenesis in the context of hepatocellular carcinoma (HCC). Methods: EVs were isolated, quantified and characterized from patients’ sera. EVs were tested vigorously, both in vitro and in vivo, by various functional assays. Proteomic analysis was performed to identify the functional components of EVs. The expression level of polymeric immunoglobulin receptor (pIgR) in circulating EVs, tumor and non-tumorous tissues of HCC patients was determined by ELISA, immunoblotting, immunohistochemistry and quantitative PCR. The functional role and underlying mechanism of EVs with an enhanced pIgR expression was elucidated. Blockage of EV-pIgR with neutralizing antibody was performed in nude mice implanted with patient-derived tumor xenograft (PDTX). Results: Circulating EVs of late-stage HCC (L-HCC) patient had significantly elevated pIgR expression when compared to the EVs released by control individuals. The augmenting effect of L-HCC patient in cancer stemness and tumorigenesis was hindered by anti-pIgR antibody. EVs enriched with pIgR consistently promoted cancer stemness and cancerous phenotypes in the recipient cells. Mechanistically, EV-pIgR-induced cancer aggressiveness was abrogated by Akt and β-catenin inhibitors, ascertaining the role of EV-pIgR through the activation of PDK1/Akt/GSK3β/β-catenin signaling axis. Furthermore, anti-pIgR neutralizing antibody attenuated the tumor growth in mice implanted with PDTX. Conclusion: The study illustrates an unrevealed role of EV-pIgR in regulating cancer stemness and aggressiveness, in which EV-pIgR activates PDK1/Akt/GSK3β/β-catenin signaling cascades. The blockage of intercellular communications mediated by EV-pIgR in the tumor microenvironment may provide a new therapeutic strategy for cancer patients.