Nanoscale analysis of prostate cancer tissue microarrays

Turiák, Lilla and Révész, Ágnes and Ács, András and Vékey, Károly and Drahos, László (2017) Nanoscale analysis of prostate cancer tissue microarrays. In: 7th BBBB International Conference on Pharmaceutical Sciences, 2017. október 5-7, Balatonfüred.

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Abstract

Cancer is the leading cause of death in the developed countries. Detecting alterations at the molecular level linked to cancer is a key aspect in developing new strategies for treatment and for diagnosis. For this purpose tissue biopsies are analyzed using various analytical techniques, such as mass spectrometry, and may result in the identification of novel biomarkers. Tissue microarrays (TMAs) contain a large number of biopsies prepared in an array format. The use of TMAs in biomedical workflows is significantly increasing in the past few years; the main challenge is the limited amount of sample available (1.5 mm diameter cores). The aim of our work is to apply advanced nanoLC-MS(MS) techniques to reliably identify various glycans and proteins from histological tissue surfaces and to apply the workflow for the analysis of prostate cancer TMAs. Integrating proteomics data with glycan analysis has recently become widespread. Glycosaminoglycans (GAGs) play crucial roles in cancer progression; however, their nanoscale analysis is still considered an analytical challenge compared to proteomics methods, which are relatively straightforward. Proteins and GAGs were analyzed using nanoLC-MS(MS) following enzymatic digestion and extraction from the surface of prostate cancer TMA slides. More than 500 proteins were identified and quantified using label free quantitation from the 1.5 mm diameter cores, including several proteoglycans (e.g., perlecan, versican, lumican) that had been previously implicated in prostate cancer. For the analysis of the GAG disaccharides self-packed nanoscale HILIC-WAX columns were packed and negative ionization mode was used. Strategies for the analysis of the limited amount of GAGs extracted from the surface of TMAs will be shown.

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