A proteomic approach to compare saliva from individuals with and without oral leukoplakia

• Published in: Journal of proteomics Vol. 151; pp. 43 - 52
• Main Authors: Camisasca, Danielle Resende, da Rós Gonçalves, Lorena, Soares, Márcia Regina, Sandim, Vanessa, Nogueira, Fábio César Sousa, Garcia, Carlos Henrique Saraiva, Santana, Rodrigo, de Oliveira, Silvia Paula, Buexm, Luisa Aguirre, de Faria, Paulo Antônio Silvestre, Dias, Fernando Luiz, Pereira, Denise de Abreu, Zingali, Russolina B., Alves, Gilda, Lourenço, Simone Queiroz Chaves
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 16-01-2017
• Subjects: 2D gel electrophoresis; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Case-Control Studies; Electrophoresis, Gel, Two-Dimensional; Humans; Keratin-10; Keratin-10 - analysis; Keratin-10 - isolation & purification; Leukoplakia, Oral - chemistry; MALDI-MS; Middle Aged; Oral leukoplakia; Precancerous Conditions - diagnosis; Proteome - analysis; Proteomics - methods; Saliva; Saliva - chemistry; 2D gel electrophoresis; MALDI-MS; Oral leukoplakia; Saliva; Keratin-10
• ISSN: 1874-3919; 1876-7737
• DOI: 10.1016/j.jprot.2016.07.029

Oral leukoplakia is the most common potentially malignant disorder in the oral cavity and can precede carcinoma. This study aimed to identify possible oral leukoplakia salivary biomarkers.
Unstimulated saliva was collected from participants and protein concentration was determined. Proteins were then precipitated with cold acetone and separated using 2DE over a pH range of 3–10. Spot demarcation and matching were performed and protein identification was done through MS analysis. Oral leukoplakia tissues were submitted to immunohistochemistry analysis for keratin 10 (CK10). A complementary analysis of oral leukoplakias that were not included previously was performed in addition.
226±10 spots were identified in oral leukoplakia 2DE gels, and 262±12 spots were identified in volunteers. Twenty-two spots were highly abundant in oral leukoplakias or not detected in the control group, such as apolipoprotein A1, alpha amylase, cystatins, keratin 10, and lysozyme precursor. All were identified. All oral leukoplakia cases were immunopositive for CK10, mainly in the superficial epithelial layers.
The 2DE salivary protein profiles of individuals with and without oral leukoplakia were observably different. CK10 appears to be an interesting protein and should be further studied in oral carcinogenesis.
MS-based proteomics enables large-scale analysis of proteins. Proteomics can provide detailed descriptions of proteomes of cells and tissues, including body fluids, and appears as a powerful tool to study human disorders. Saliva is readily accessible through non invasive collection and can mirror diverse disease states. Saliva from both diseased and healthy subjects can be analyzed through 2DE and differences between groups could be found. Routine immunohistochemistry analysis confirmed one of these findings, with CK10 being positive tissues from individuals with oral leukoplakia. Therefore, the present study allows insights into development of an important potential oral cancer precursor, named oral leukoplakia. However, the results can be extrapolated and tested in other precancer states, such as proliferative verrucous leukoplakia, patients at risk of oral cancer due to lifestyle behavior and/or cancer history in the family or even those who are under surveillance after a treated primary oral cancer.
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•2DE is suitable to identify differences in salivary proteins from OL and controls.•Common proteins, such as alpha amylase, immunoglobulin and albumin were identified.•Keratin 10, apolipoprotein A1, lysozyme precursor deserve further investigation.•Keratin 10 was present in all tissues from two different set of patients.•Areas with orthokeratin dysplasia were more prone to be CK10 positive.