Quantification of Pancreatic Cancer Proteome and Phosphorylome: Indicates Molecular Events Likely Contributing to Cancer and Activity of Drug Targets

被引：42

作者：

Britton, David ^{[1
]}

Zen, Yoh ^{[2
]}

Quaglia, Alberto ^{[2
]}

Selzer, Stefan ^{[1
]}

Mitra, Vikram ^{[1
]}

Loessner, Christopher ^{[1
]}

Jung, Stephan ^{[1
]}

Boehm, Gitte ^{[1
]}

Schmid, Peter ^{[1
]}

Prefot, Petra ^{[1
]}

Hoehle, Claudia ^{[1
]}

Koncarevic, Sasa ^{[1
]}

Gee, Julia ^{[4
]}

Nicholson, Robert ^{[4
]}

Ward, Malcolm ^{[1
]}

Castellano, Leandro ^{[3
]}

Stebbing, Justin ^{[3
]}

Zucht, Hans Dieter ^{[1
]}

Sarker, Debashis ^{[2
]}

Heaton, Nigel ^{[2
]}

Pike, Ian ^{[1
]}

机构：

[1] Proteome Sci Plc, Cobham, England

[2] Kings Coll Hosp London, Inst Liver Studies, London SE5 8RX, England

[3] Univ London Imperial Coll Sci Technol & Med, Fac Med, Dept Surg & Canc, London, England

[4] Cardiff Univ, Cardiff Sch Pharm & Pharmaceut Sci, Cardiff CF10 3AX, S Glam, Wales

来源：

PLOS ONE | 2014年 / 9卷 / 03期

关键词：

CELL-MIGRATION; TYROSINE PHOSPHORYLATION; HEPATOCELLULAR-CARCINOMA; COLORECTAL-CANCER; BETA-CATENIN; KINASE; EXPRESSION; GENE; GROWTH; DOMAIN;

D O I：

10.1371/journal.pone.0090948

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Objective: LC-MS/MS phospho-proteomics is an essential technology to help unravel the complex molecular events that lead to and propagate cancer. We have developed a global phospho-proteomic workflow to determine activity of signaling pathways and drug targets in pancreatic cancer tissue for clinical application. Methods: Peptides resulting from tryptic digestion of proteins extracted from frozen tissue of pancreatic ductal adenocarcinoma and background pancreas (n = 12), were labelled with tandem mass tags (TMT 8-plex), separated by strong cation exchange chromatography, then were analysed by LC-MS/MS directly or first enriched for phosphopeptides using IMAC and TiO2, prior to analysis. In-house, commercial and freeware bioinformatic platforms were used to identify relevant biological events from the complex dataset. Results: Of 2,101 proteins identified, 152 demonstrated significant difference in abundance between tumor and non-tumor tissue. They included proteins that are known to be up-regulated in pancreatic cancer (e.g. Mucin-1), but the majority were new candidate markers such as HIPK1 & MLCK. Of the 6,543 unique phosphopeptides identified (6,284 unique phosphorylation sites), 635 showed significant regulation, particularly those from proteins involved in cell migration (Rho guanine nucleotide exchange factors & MRCK alpha) and formation of focal adhesions. Activator phosphorylation sites on FYN, AKT1, ERK2, HDAC1 and other drug targets were found to be highly modulated (>= 2 fold) in different cases highlighting their predictive power. Conclusion: Here we provided critical information enabling us to identify the common and unique molecular events likely contributing to cancer in each case. Such information may be used to help predict more bespoke therapy suitable for an individual case.

引用

页数：20

共 54 条

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