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. 2017 May-Jun;14(3):197-211.
doi: 10.21873/cgp.20031.

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

Gurler Akpinar  1   2 , Murat Kasap  3   2 , Nuh Zafer Canturk  4 , Mehin Zulfigarova  3   2 , Eylül Ece Islek  3   2 , Sertac Ata Guler  4 , Turgay Simsek  4 , Zeynep Canturk  5
Affiliations

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

Gurler Akpinar et al. Cancer Genomics Proteomics. 2017 May-Jun.

Abstract

Background/aim: To unveil the pathophysiology of primary hyperparathyroidism, molecular details of parathyroid hyperplasia and adenoma have to be revealed. Such details will provide the tools necessary for differentiation of these two look-alike diseases. Therefore, in the present study, a comparative proteomic study using postoperative tissue samples from the parathyroid adenoma and parathyroid hyperplasia patients was performed.

Materials and methods: Protein extracts were prepared from tissue samples (n=8 per group). Protein pools were created for each group and subjected to DIGE and conventional 2DE. Following image analysis, spots representing the differentially regulated proteins were excised from the and used for identification via MALDI-TOF/TOF analysis.

Results: The identities of 40 differentially-expressed proteins were revealed. Fourteen of these proteins were over-expressed in the hyperplasia while 26 of them were over-expressed in the adenoma.

Conclusion: Most proteins found to be over-expressed in the hyperplasia samples were mitochondrial, underlying the importance of the mitochondrial activity as a potential biomarker for differentiation of parathyroid hyperplasia from adenoma.

Keywords: 2DE; MALDI-TOF/TOF; Parathyroid adenoma; Parathyroid gland; Parathyroid hyperplasia; proteomics.

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Figures

Figure 1
Figure 1. Summary of the experimental approaches used in this study. (A) The gel images from the Coomassie-stained and DIGE experiments. (B) The in silico produced master gel images for comparison of 2DE vs DIGE experiments. (C) Venn diagram depicting common protein spots for the 2DE and DIGE gels. The common spots were identified by MALDI-TOF/TOF analysis
Figure 2
Figure 2. Enlarged images of the selected 2D gel spots to highlight the differential expression. The percent volumes of the identified proteins in PA and PH groups. The values for the bar graphs from the comperative 2DE gel analysis experiment via PDQuest Advance software
Figure 3
Figure 3. Representative 2DE maps of PH (A) and PA (B) sample proteins. Spot numbers (SSP numbers) indicate the proteins identified by MALDI TOF/TOF MS and refer to the numbers reported in Table IV
Figure 4
Figure 4. Functional and localization analysis of the up-regulated proteins. (A) Pie charts representing functional categorization of the identified up-regulated proteins for the PA and the PH groups. (B) Pie charts representing cellular localization for the identified up-regulated proteins for the PA and the PH groups
Figure 5
Figure 5. Western blot analysis of protein pools from the PA and the PH groups. An antibody mixture, OXPHOS, was used to blot the membrane. β-Actin was used as an internal control
Figure 6
Figure 6. The identified proteins in the PA (A) and the PH (B) groups were mapped by searching the STRING v10.0 software with a confidence level of 0.4. Colored lines between the proteins indicate the various types of interaction evidence
Figure 7
Figure 7. Validation of the regulations observed in LMNA, CH60, HNRNP and UCHL-1 by western blotting. β-Actin was used as an internal control. The bar graphs were created from densitometric analysis data which was performed by using the QuantityOne software. To represent each group (PA and PH) four wells were loaded and standard deviation was calculated based on intensities of the bands of each well
Figure 8
Figure 8. Representative gel images of DIGE experiments
See this image and copyright information in PMC

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