Background Ameloblastoma (AME) is characterized by a locally invasive growth pattern. In an attempt to justify the aggressiveness of neoplasms, the investigation of the role of stem cells has gained prominence. The SOX-2, NANOG and OCT4 proteins are important stem cell biomarkers. Methodology To verify the expression of these proteins in tissue samples of AME, dentigerous cyst (DC) and dental follicle (DF), immunohistochemistry was performed and indirect immunofluorescence were performed on the human AME (AME-hTERT) cell line. Results Revealed expression of SOX-2, NANOG and OCT4 in the tissue samples and AME-hTERT lineage. Greater immunostaining of the studied proteins was observed in AME compared to DC and DF (p < 0.001). Conclusions The presence of biomarkers indicates a probable role of stem cells in the genesis and progression of AME.

Objective The aim of this study was to evaluate and compare alterations in gene expression using two distinct immortalization methods (hTERT and HPV16‐E6/E7) in ameloblastoma cell lines. Materials and Methods A primary cell culture derived from human ameloblastoma (AME‐1) was established and immortalized by two different methods using a transfection processes to hTERT and HPV‐E6/E7. The RNA‐seq was used to verify which immortalization method had less influence on gene expression. It was performed in four steps: extraction and collection of mRNA, PCR amplification, comparison with the human reference genome, and analysis of differential expression. The genes with differentiated expression were identified and mapped. Results RNA‐seq revealed genetic alterations in ameloblastoma cell lines after the immortalization process, including increased expression of tumor genes like MYC, E2F1, BRAF, HRAS, and HTERT, and a decrease in tumor suppressor genes like P53, P21, and Rb. Conclusions It is possible to affirm that cell immortalization is not an inert method regarding gene regulation mechanisms and the hTERT method (AME‐TERT) presented fewer changes in gene expression levels.

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