Evaluation of IBSP, BGLAP, and NFIC gene and protein expression in primordial odontogenic tumors: preliminary results

Abstract

Objective: To evaluate the gene expression of IBSP, BGLAP, and NFIC, and to determine the immunoexpression of the BGLAP and NFIC proteins in Primordial Odontogenic Tumor (POT), with the aim of improving the understanding of its pathogenesis.

Methodos: Five formalin-fixed, paraffin-embedded POT samples were processed for reverse transcription quantitative polymerase chain reaction (RT-qPCR) and immunohistochemistry (IHC) analyses. For RT-qPCR, total RNA was extracted, complementary DNA (cDNA) was synthesized, and the expression levels of IBSP, BGLAP, and NFIC were assessed. Relative gene expression was quantified using the 2(-ΔΔCt) method. For immunohistochemistry, 5-µm tissue sections were obtained, deparaffinized, rehydrated, and subjected to antigen retrieval. Sections were incubated with primary antibodies against BGLAP or NFIC, followed by incubation with a biotinylated anti-mouse/anti-rabbit secondary antibody and a streptavidin–peroxidase complex. Immunoreactivity was visualized using 3,3′-diaminobenzidine (DAB), and the sections were counterstained with Mayer’s hematoxylin.

Results: POT samples exhibited low expression levels of NFIC and IBSP, while BGLAP expression was undetectable at the gene level. Immunohistochemical analysis revealed BGLAP immunoreactivity in the epithelial component (nucleus and cytoplasm) as well as in connective tissue cells, except in one case. NFIC protein expression was not detected in either epithelial or connective tissue cells, except in one case.

Conclusions: These findings suggest that POT is characterized by significant disruption of molecular pathways associated with odontoblastic differentiation and dentin formation, supporting the hypothesis that this neoplasm represents a developmental arrest at very early stages of odontogenesis.