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

Abstract

BACKGROUND:

Proteomic 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.

RESULTS:

The 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.

CONCLUSIONS:

In 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.

• Authors: Elena Donadio, Laura Giusti, Filomena Cetani, Ylenia Da Valle, Federica Ciregia, Gino Giannaccini, Elena Pardi, Federica Saponaro, Liborio Torregrossa, Fulvio Basolo, Claudio Marcocci, Antonio Lucacchini
• Journal: Proteome science (Proteome Sci) Vol. 9 Issue 1 Pg. 29 (Jun 08 2011) ISSN: 1477-5956 [Electronic] England
• PMID: 21651755 (Publication Type: Journal Article)