Establishment and use of a three-dimensional ameloblastoma culture model to study the effects of butyric acid on the transcription of growth factors and laminin β3

Abstract Objective This study aimed to establish a three-dimensional (3D) culture method for ameloblastoma cell lines and to use the model to investigate the effect of butyric acid (BA), a periodontopathic bacterial metabolite, on the malignant transformation of ameloblastoma. Design Three ameloblastoma cell lines (HAM1, HAM2, and HAM3) established from the same tumor were used in this study. A 3D culture model was established in low absorption dishes and was incubated for 48 h. The effects of BA on the transcription of growth factors and LMβ3 were examined by real-time reverse transcription PCR. Various BA concentrations (0.02, 0.2, 2, and 20 mM) were used to stimulate the cell cultures for 6 and 12 h. Results A 3D culture model was established. Gene expression levels of epithelial growth factor (EGF), transforming growth factor beta 1 (TGFβ1), and laminin β3 (LMβ3) were higher in 3D than in 2D cultures. Cell morphology in 3D cultures did not change, while the transcription levels of EGF, TGFβ1, and LMβ3 were upregulated by BA in all cell lines. Conclusion The 3D culture model is more responsive to BA than the 2D culture model, and there is a possibility that the malignancy and progression of ameloblastoma via laminin 332 (LM332) is mediated by BA.