Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2025 Mar;31(3):865-878.
doi: 10.1111/odi.15109. Epub 2024 Aug 18.

Pleomorphic adenoma and carcinoma ex-pleomorphic adenoma tumorigenesis: A proteomic analysis

Virgílio Gonzales Zanella  1   2 , Sara Ferreira Dos Santos Costa  3 , Lauren Frenzel Schuch  4 , Emily Ferreira Salles Pilar  5 , Adriana Franco Paes Leme  4 , Jean Nunes Dos Santos  6 , Syed Ali Khurram  7 , Fatima Elalawy  7 , Lynne Bingle  7 , Fabio Daumas Nunes  8 , Felipe Paiva Fonseca  3   4 , Pablo Agustin Vargas  4 , Manoela Domingues Martins  1   4 , Vivian Petersen Wagner  4   7   8
Affiliations

Pleomorphic adenoma and carcinoma ex-pleomorphic adenoma tumorigenesis: A proteomic analysis

Virgílio Gonzales Zanella et al. Oral Dis. 2025 Mar.

Abstract

Objectives: To conduct a comprehensive proteomic analysis of normal salivary gland tissue, pleomorphic adenoma (PA), and carcinoma ex-pleomorphic adenoma (CXPA), and validate the proteomic findings using immunohistochemistry.

Methods: Six normal salivary gland tissues, seven PA and seven CXPA samples underwent laser microdissection followed by liquid chromatography coupled to mass spectrometry. Protein identification and quantification were performed using MaxQuant software. Statistical analysis and functional enrichment were conducted using the Perseus platform and STRING tool, respectively. Immunohistochemistry was used for validation.

Results: Comparative proteomic analysis revealed 2680 proteins across the three tissue types, with 799 significantly altered between groups. Translocation protein SEC63 homolog, Annexin A6 and Biglycan were up-regulated in CXPA compared to PA. Decorin was markedly up-regulated in both PA and CXPA compared to normal salivary gland (log2 fold changes of 7.58 and 7.38, respectively). Validation confirmed elevated levels of Biglycan and Decorin in the extracellular matrix of CXPA compared to PA.

Conclusions: Proteomic analysis identified differential protein expression patterns associated with malignant transformation of PA into CXPA. Findings indicate a crucial role for extracellular matrix proteins, specifically Biglycan and Decorin, in the tumorigenic progression of PA and CXPA.

Keywords: carcinoma ex‐pleomorphic adenoma; glandular and epithelial; head and neck neoplasms; proteomic; rare diseases.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

FIGURE 1
FIGURE 1
Proteins identified and quantified in the three groups. (a) Venn diagram of common and unique proteins in the three groups showed 2347 proteins identified in common in the three groups analyzed, 168 proteins shared between neoplasm groups, 58 proteins common between CA ex‐PA and NSG, and 19 proteins shared between PA and NGS. Of the total number of proteins identified, only 62, 22, and 4 were exclusively identified in PA, CA ex‐PA, and NSG, respectively. (b) In an exploratory analysis using a hierarchical method, the dendrogram of the proteins identified showed the hierarchy and clusters between the samples in each group. The cases analyzed are listed on the right vertical axis, whereas the horizontal axis shows the distance between the clusters when they are joined. The result indicated the initial formation of two clusters: The neoplasm cluster and the NSG cluster, with the CA ex‐PA and PA clusters being more closely correlated with each other and different from the NGS cluster. In addition, there was greater similarity between the samples in each group, demonstrated by the high correlation between cases in the same group. (c) The heat map was used to compare the abundance of proteins between the samples analyzed in the three groups (Z‐score log2 intensity values). Each row represents an identified protein and each column is a single sample analyzed. The intensity of the color at each intersection point indicates the level of abundance of each protein per sample, with high abundance shown in red and low abundance in blue. The cluster analysis clearly identified the main groupings as shown in the color bar at the top in green, red, and blue representing the NSG, PA, and CXPA groups, respectively.
FIGURE 2
FIGURE 2
(a) Principal component analysis (PCA) presented as a score‐plot 2D of the main components observed among the three groups demonstrated the separation between samples according to abundance variation. The colored dots represent a single sample in each group. The samples in each group are close to each other forming three distinct groups. (b) Protein–protein interactions (PPI) networking showing groups of highly correlated proteins as indicated by the black traces (the thicker the trace, the stronger the correlation between the proteins).
FIGURE 3
FIGURE 3
Representative images of low‐power and high‐power view of (a) Biglycan and (b) Decorin expression in pleomorphic adenoma and carcinoma ex‐pleomorphic adenoma. Statistical analysis confirmed that expression of (c) Biglycan and (d) decorin were higher in CXPA compared to PA (Mann–Whitney U‐test, *p < 0.05; **p < 0.01).
FIGURE 4
FIGURE 4
Representative image of Biglycan pattern of expression within one case of CXPA. Note that residual areas of PA showed absence of expression in the ECM component while areas of malignant transformation (characterized by bizarre pleomorphic cells) showed intense expression.
See this image and copyright information in PMC

References

    1. Abiko, Y. , Nishimura, M. , Mitamura, J. , Rahemtulla, F. , Scott, P. G. , Dodd, C. M. , Mizoguchi, I. , & Kaku, T. (1999). An immunohistochemical study of the localization of biglycan, decorin and large chondroitin‐sulfate proteoglycan in pleomorphic adenoma. Acta Histochemica et Cytochemica, 32(6), 459–463.
    1. Altemani, A. , Martins, M. T. , Freitas, L. , Soares, F. , Araújo, N. S. , & Araújo, V. C. (2005). Carcinoma ex pleomorphic adenoma (CXPA): Immunoprofile of the cells involved in carcinomatous progression. Histopathology, 46(6), 635–641. - PubMed
    1. Andreasen, S. , Therkildsen, M. H. , Bjørndal, K. , & Homøe, P. (2016). Pleomorphic adenoma of the parotid gland 1985‐2010: A Danish nationwide study of incidence, recurrence rate and malignant transformation. Head & Neck, 38(Suppl 1), E1364–E1369. - PubMed
    1. Bankhead, P. , Loughrey, M. B. , Fernández, J. A. , Dombrowski, Y. , McArt, D. G. , Dunne, P. D. , McQuaid, S. , Gray, R. T. , Murray, L. J. , Coleman, H. G. , James, J. A. , Salto‐Tellez, M. , & Hamilton, P. W. (2017). QuPath: Open‐source software for digital pathology image analysis. Scientific Reports, 7(1), 16878. - PMC - PubMed
    1. Carnielli, C. M. , Macedo, C. C. S. , De Rossi, T. , Granato, D. C. , Rivera, C. , Domingues, R. R. , Pauletti, B. A. , Yokoo, S. , Heberle, H. , Busso‐Lopes, A. F. , Cervigne, N. K. , Sawazaki‐Calone, I. , Meirelles, G. V. , Marchi, F. A. , Telles, G. P. , Minghim, R. , Ribeiro, A. C. P. , Brandão, T. B. , de Castro, G., Jr. , … Paes Leme, A. F. (2018). Combining discovery and targeted proteomics reveals a prognostic signature in oral cancer. Nature Communications, 9(1), 3598. - PMC - PubMed