Proteomics and N-glycosylation analysis of prostate cancer biopsies

Sugár, Simon and Tóth, Gábor and Bugyi, Fanni and Vékey, Károly and Drahos, László and Turiák, Lilla (2021) Proteomics and N-glycosylation analysis of prostate cancer biopsies. In: MSACL 2021 EU, 2021.08.31-09.03, online.

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Abstract

Introduction Analysis of tissue specimen using mass spectrometry offers valuable information of biological processes taking place at the origin of a disease. Identifying differences in protein expression levels and site-specific N-glycosylation of glycoproteins among cancerous and healthy tissues can be an attractive approach in biomarker research. Our aim was to analyze prostate cancer (PCa) tissue microarray samples (n=95) and compare protein expression levels and changes in N-glycosylation features among various pathological grades of PCa and healthy tissues. Methods Following on surface proteolytic digestion [1] glycopeptides were enriched using acetone precipitation. Peptide and glycopeptide fractions were analyzed separately using reversed phase nanoHPLC-MS(MS) and nanoHPLC-MS. Label free protein quantitation was performed using MaxQuant, while glycopeptides were identified and quantified using Byonic and Glycopattern software, respectively. Statistical analysis was carried out using Perseus. Results Several statistically significant proteins were identified that were altered between different cancer grades and healthy tissue. STRING analysis revealed protein pathways involved in disease progression. Regarding N-glycosylation analysis the changes identified at specific glycosylation sites were of interest. Alterations in sialylation, fucosylation and galactosylation of individual glycosites were detected that could potentially be important targets of future studies. We found that in case of Collagen 6 subunit both protein expression levels and fucosylation at N785 changed significantly between the different pathological grades and healthy tissue. Conclusions Our integrated proteomics and glycoproteomics workflow was used to identify molecular alterations occurring during PCa progression from limited size tissue samples and can also be used to analyze other types of cancerous tissue biopsies in the future.

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