A proteomic approach to study parathyroid glands

Abstract

Parathyroid tumours are heterogeneous and in some cases the diagnosis may be difficult using histological features. In this study we used a two-dimensional electrophoresis (2D)/mass spectrometry (MS)-based approach to examine the global changes of parathyroid adenoma tissues protein profile compared to the parathyroid normal tissues. Validation of protein expression was performed by immunoblotting using specific antibodies. Ingenuity software was used to identify the biological processes to which these proteins belong and to construct a potential network. A total of 30 proteins were found to be differentially expressed, of which 22 resulted in being over-expressed. Proteins identified by 2D/MS/MS proteomics were classified into functional categories and a major change (≥ 2-fold) in terms of expression was found in proteins involved in response to biotic stimuli, cell organization and signal transduction. After Ingenuity analysis, 14-3-3 ζ/δ appears to be a key protein in the network of parathyroid adenoma, where it is linked to other proteins such as annexin A2, B box and SPRY domain-containing protein (BSPRY), p53 and epidermal growth factor receptor (EGFR). Our results suggest that the proteomic approach was able to differentiate the protein profiles of normal parathyroid and parathyroid adenoma and identify a panel of proteins which are differentially expressed. The functional role of these proteins in the network of intracellular pathways is discussed.

Proteomics Analysis of Tissue Samples Reveals Changes in Mitochondrial Protein Levels in Parathyroid Hyperplasia over Adenoma

“…In order to make comparisons for the regulated proteins, a Venn diagram was created using the data provided by the earlier aforementioned studies and our data. ANXA2 and LMNA reported by Guisti et al, (12) and ATP5H reported by Varshney et al, (13) were also detected in our study.…”

“…Bioinformatic analysis of the relevant pathways based on the regulated proteins predicted that 14-3-3 ζ/δ is the key protein in PA network. The study by Giusti et al,(12) demonstrated the use of a proteomic approach for the discovery of potential markers in differential diagnosis of PA. On this line of study, the later work by Varshney et al, (13) monitored the changes in parathyroid proteome in patients with sporadic parathyroid adenoma in comparison to normal parathyroid tissue. Fifteen differentially expressed proteins were reported.…”

“…In addition to the genetic studies, proteomic studies are also needed. There are two major studies evaluated the changes at the proteome level in comparison to healthy controls in PA (12,13). A list of differentially-regulated proteins generated by these studies is useful to reveal the metabolic pathways that may play roles in transformation to PA.…”

Primary hyperparathyroidism

“…Interestingly, mutations were germline in two patients with CDKN1B mutations and in one carrying a mutation in AIP Contradictory results have been reported on the presence of CTNNB1 (encoding β-catenin) mutations involving serine/threonine phosphorylation 86,87 . Up to 30 proteins, involved in various cellular pathways (for example, response to biotic stimuli, cell organization and signal transduction), are differentially expressed in normal parathyroid and parathyroid adenomas 88 . Of these various proteins, 14-3-3ζ/ δ may be one of the key proteins in the pathogenesis of parathyroid adenoma.…”

Molecular Alterations in Sporadic Primary Hyperparathyroidism

“…Giusti and collaborators carried out comparative analysis to examine the changes in protein profile between adenomas and normal parathyroid tissue [77]. These authors verified the presence of 30 deregulated proteins in parathyroid adenomas, 22 of them being overexpressed when compared to normal parathyroid tissue [77].…”