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Phosphopeptide Enrichment Coupled with Label-free Quantitative Mass Spectrometry to Investigate the Phosphoproteome in Prostate Cancer
Published on: August 2, 2018
Evaluation of glycophenotype in prostatic neoplasm by chemiluminescent assay
Lúcia Patrícia Bezerra Gomes da Silva1, Sinara Mônica Vitalino de Almeida2, Luiza Rayanna Amorim de Lima1
1Departamento de Bioquímica and Laboratório de Imunopatologia Keizo Asami (LIKA), Universidade Federal de Pernambuco, Cidade Universitária Recife, Pernambuco, Brasil.
Abstract:
This work aimed to evaluate the glycophenotype in normal prostate, bening prostatic hyperplasia (BPH) and prostatic adenocarcinoma (PCa) tissues by a chemiluminescent method. Concanavalin A (Con A), Ulex europaeus agglutinin (UEA-I) and Peanut agglutinin (PNA) lectins were conjugated to acridinium ester (lectins-AE). These conjugates remained capable to recognize their specific carbohydrates. Tissue samples were incubated with lectins-AE. The chemiluminescence of the tissue-lectin-AE complex was expressed in relative light units (RLU). Transformed tissues (0.25 cm(2) by 8 µm of thickness) showed statistical significant lower α-D-glucose/mannose (BPH: 226,931 ± 17,436; PCa: 239,520 ± 12,398) and Gal-β(1-3)-GalNAc (BPH: 28,754 ± 2,157; PCa: 16,728 ± 1,204) expression than normal tissues (367,566 ± 48,550 and 409,289 ± 22,336, respectively). However, higher α-L-fucose expression was observed in PCa (251,118 ± 14,193) in relation to normal (200,979 ± 21,318) and BHP (169,758 ± 10,264) tissues. It was observed an expressive decreasing of the values of RLU by inhibition of the interaction between tissues and lectins-AE using their specific carbohydrates. The relationship between RLU and tissue area showed a linear correlation for all lectin-AE in both transformed tissues. These results indicated that the used method is an efficient tool for specific, sensitive and quantitative analyses of prostatic glycophenotype.
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