The aim of this study was to investigate alterations in the EGFR gene and its protein expression for a better understanding of the biologic behavior of ameloblastoma.Twenty-five samples of ameloblastoma were selected, and dual-color fluorescence in situ hybridization assay was performed. The results of the assay and immunohistochemistry reaction for EGFR and Ki67 were associated with clinicopathologic features and recurrence.All analyzed cases presented disomy without any gene polysomy or amplification. With regard to EGFR immunoexpression, 3 cases (12%) were considered negative, and 22 (88%) were positive, of which 13 (52%) were weak and 9 (36%) were strong. All samples presented low positivity for Ki67. There was no association between EGFR expression and clinicopathologic features or recurrence (P > .05). In some cases, EGFR immunoexpression was observed without gene amplification.Ameloblastoma development, progression, or recurrence does not appear to be related to EGFR amplification or polysomy.