In oral squamous cell carcinoma (OSCC), involvement and activation of the Hedgehog pathway (HH) may be related to epithelial-mesenchymal transition and cell proliferation. The present study aimed to evaluate epithelial-mesenchymal transition and proliferative potential in OSCC cases demonstrating activation of the HH pathway. Twenty-three GLi-1-positive OSCC cases were submitted to immunohistochemical detection of Snail, Slug, N-cadherin, E-cadherin, β-catenin, and MCM3 proteins. Clinicalpathologic immunoexpression data were obtained from the invasion front and tumor islets, and then compared. At the invasion front, OSCC cases presented positive Snail, Slug, and MCM3 expression in the nuclei of tumor cells. Loss of membrane and cytoplasmic expression of E-cadherin and β-catenin was also observed. Positive N-cadherin expression was observed in 31.78% of the cases. GLi-1 immunoexpression was associated with loss of membrane E-cadherin (P<0.001), membrane β-catenin (P<0.001), and cytoplasmic β-catenin (P=0.02) expression. In the tumor islets, we observed nuclear expression of GLi-1, Snail, Slug, and MCM3. E-cadherin and β-catenin showed positivity in tumor cell membranes. Statistically significant positive correlations between GLi-1 and Snail (P=0.05), E-cadherin (P=0.01), and cytoplasmic β-catenin (P=0.04) were found. GLi-1 was associated with clinical staging, while membrane β-catenin expression was related to the presence of metastasis in lymph nodes and to clinical staging. The HH pathway may be involved in regulating the expression of the mesenchymal phenotype. The loss of membrane E-cadherin and β-catenin expression was observed at the tumor front region, whereas cell adhesion protein expression was detected in tumor islets regardless of MCM3.