Upregulation of tropomyosin alpha-4 chain in patients’ saliva with oral squamous cell carcinoma as demonstrated by Phage display

• Published in: Scientific reports Vol. 9; no. 1; pp. 18399 - 8
• Main Authors: Faria, Paula Cristina Batista, Carneiro, Ana Paula, Binato, Renata, Nascimento, Rafael, Santos, Paula Souza, Fagundes, Deborah, da Silva, Sindeval José, Loyola, Adriano Mota, Abdelhay, Eliana, Goulart, Luiz Ricardo
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 05.12.2019; Nature Publishing Group
• Subjects: 13/51; 38/1; 38/22; 38/23; 631/61/185; 631/67/1665; 82/58; Actin; Adult; Aged; Aged, 80 and over; Amino Acid Sequence; Antibodies; Antigens; Binding sites; Bioinformatics; Biomarkers; Carcinoma, Squamous Cell - diagnosis; Carcinoma, Squamous Cell - genetics; Carcinoma, Squamous Cell - metabolism; Carcinoma, Squamous Cell - pathology; Cloning; Computational Biology - methods; Connective tissue; Connective Tissue - metabolism; Connective Tissue - pathology; Connective tissues; Cytoskeleton; Discovery tools; Enzyme-Linked Immunosorbent Assay; Epithelial cells; Epithelial Cells - metabolism; Epithelial Cells - pathology; Female; Filaments; Gel electrophoresis; Gene Expression; Humanities and Social Sciences; Humans; Invasiveness; Keratin; Laboratories; Ligands; Longitudinal studies; Male; Mass spectrometry; Mass spectroscopy; Middle Aged; Models, Molecular; Mortality; Mouth Neoplasms - diagnosis; Mouth Neoplasms - genetics; Mouth Neoplasms - metabolism; Mouth Neoplasms - pathology; multidisciplinary; Neoplasm Proteins - genetics; Neoplasm Proteins - metabolism; Oral cancer; Oral squamous cell carcinoma; Peptide Library; Phage display; Polyacrylamide; Protein Structure, Secondary; Proteins; Proteomes; Saliva; Saliva - chemistry; Salivary gland; Salivary Glands - metabolism; Salivary Glands - pathology; Science; Science (multidisciplinary); Scientific imaging; Sensitivity and Specificity; Single-Chain Antibodies - chemistry; Squamous cell carcinoma; Tropomyosin; Tropomyosin - genetics; Tropomyosin - metabolism; Tumor necrosis factor-TNF; Up-Regulation
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-019-54686-x

Patients with oral squamous cell carcinoma (OSCC) present significant alterations in their saliva proteome. We have used the shotgun Phage Display (PD) technology to identify candidate proteins that were upregulated in saliva of OSCC by selecting ligands to salivary proteins from a single-chain variable fragment (scFv) PD combinatorial library. After two selection cycles, the highly reactive clone scFv-D09 was able to distinguish saliva of OSCC patients from healthy subjects by enzyme-linked immunosorbent assay (ELISA) with sensitivity and specificity of 96.67%. Additionally, the scFv-D09 clone presented a positive immunostaining for invasive malignant epithelial cells in the connective tissue, keratin pearls in the OSCC, and ducts of salivary glands. We have further identified the target protein as the tropomyosin alpha-4 chain (TPM4) by two-dimensional polyacrylamide gel electrophoresis and mass spectrometry, and its binding to the scFV-D09 was demonstrated by bioinformatics. Briefly, we have identified TPM4 as upregulated salivary protein in patients with OSCC, which plays a central role in stabilizing cytoskeleton actin filaments, probably linked with tumor tissue remodeling. Long-term longitudinal studies are needed to validate TPM4 as a potential marker of a malignant process.