Evaluation of formalin-fixed paraffin-embedded tissues in the proteomic analysis of parathyroid glands

Proteome Science

Research

Filomena Cetani¹  Federica Saponaro¹  Claudio Marcocci¹  Elena Pardi¹  Ylenia Da Valle²  Elena Donadio²  Gino Giannaccini²  Antonio Lucacchini²  Federica Ciregia²  Laura Giusti²  Liborio Torregrossa³  Fulvio Basolo³ 

[1] Department of Endocrinology and Metabolism, Orthopedics and Traumatology, Occupational Medicine University of Pisa, Via Paradisa 2, 56124, Pisa, Italy;Department of Psychiatry, Neurobiology, Pharmacology and Biotechnology, University of Pisa, Via Bonnanno 6, 56126, Pisa, Italy;Department of Surgery, University of Pisa, Via Paradisa 2, 56124, Pisa, Italy;

关键词: Sodium Dodecyl Sulfate;    Parathyroid Adenoma;    Ammonium Bicarbonate;    Parathyroid Carcinoma;    Parathyroid Tissue;   

DOI  :  10.1186/1477-5956-9-29

 received in 2011-02-17, accepted in 2011-06-08,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundProteomic research in the field of parathyroid tissues is limited by the very small dimension of the glands and by the low incidence of cancer lesions (1%). Formalin-fixed paraffin-embedded (FFPE) tissue specimens are a potentially valuable resource for discovering protein cancer biomarkers. In this study we have verified the applicability of a heat induced protein extraction from FFPE parathyroid adenoma tissues followed by a gel-based or gel-free proteomic approach in order to achieve protein separation and identification.ResultsThe best results for high quality MS spectra and parameters, were obtained by using a gel-free approach, and up to 163 unique proteins were identified. Similar results were obtained by applying both SDS-out and SDS-out + TCA/Acetone techniques during the gel-free method. Western blot analysis carried out with specific antibodies suggested that the antigenicity was not always preserved, while specific immunoreactions were detected for calmodulin, B box and SPRY domain-containing protein (BSPRY), peroxiredoxin 6 (PRDX 6) and parvalbumin.ConclusionsIn spite of some limitations mainly due to the extensive formalin-induced covalent cross-linking, our results essentially suggest the applicability of a proteomic approach to FFPE parathyroid specimens. From our point of view, FFPE extracts might be an alternative source, especially in the validation phase of protein biomarkers when a large cohort of samples is required and the low availability of frozen tissues might be constraining.

【 授权许可】

Unknown   
© Donadio et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.