Integrated Proteomics and Glycomics of Prostate Cancer Tissue Microarrays

Turiák, Lilla and Tóth, Gábor and Ács, András and Révész, Ágnes and Vékey, Károly and Drahos, László (2017) Integrated Proteomics and Glycomics of Prostate Cancer Tissue Microarrays. In: 11th Balaton Symposium on high-performance separation methods, 2017. szeptember 6-8, Siófok.

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Abstract

Prostate cancer is amongst the most common types of cancer in men. The current screening method - which is based on the measurement of Prostate Specific Antigen (PSA) level in the blood - is still controversial. Tissue biopsies are often used in mass spectrometry based biomarker research and hold great promise in understanding biochemical mechanisms underlying diseases such as cancer. Detecting anomalies at the molecular level are necessary for early stage diagnosis and successful treatment. Tissue microarrays (TMA) are a pathologically well characterized source of biopsies fixed on a tissue slide and arranged in an array format. In the past they have been predominantly characterized by immunohistochemistry, while LC-MS based analysis of various compound classes e.g. proteins, glycans etc. are still in the early stages due to several difficulties related to the limited amount of sample (1.5 mm diameter cores). The aim of our work is to apply advanced nanoLC-MS(MS) techniques to reliably identify glycosaminoglycans (GAGs) and proteins from the surfaces of prostate cancer TMAs. Integrating proteomics data with glycan analysis is recently gaining attention. GAGs are known to play critical roles in cancer progression; however, their nanoscale analysis is still considered an analytical challenge compared to proteomics methods, where sample preparation and nanoLC-MS(MS) methods can be considered more or less straightforward. Proteins and GAGs (heparan sulfate and chondroitin sulfate) from the same TMA core were analyzed using separate nanoLC-MS(MS) methodologies following enzymatic digestion and extraction from the surface of a prostate cancer TMA slide containing 24 cores of adenocarcinoma sample. Analysis and separation of the limited amount of GAG disaccharides requires use of self-packed nanoscale HILIC-WAX columns and negative ionization mode; complications and strategies for their analysis will be shown. More than 500 proteins were identified and quantified using label free quantitation from the 1.5 mm diameter TMA cores, amongst them several proteoglycans (e.g., perlecan, versican, lumican) that had been previously implicated in prostate cancer. Proteoglycan content varied in different stages of cancer and may be a potentially useful indicator of prostate cancer progression.

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