The antitumor activity of Genipa americana bark lectin (GaBL) was evaluated for the first time against cell lines of human skin cancer (A431), melanoma (B16), and squamous cell carcinoma of the tongue (SCC9). Cancer cell lines were treated with 10 µg/ml of GaBL to assess cell viability, cell migration and invasion, as well as the identification of cell membrane alterations associated with apoptosis. Real-time polymerase chain reaction for caspase-3 was performed to verify if apoptosis is activated by lectin treatment. The mRNA expression of proteins (E-cadherin, type I collagen) related to the epithelial-mesenchymal transition was also analyzed. GaBL decreased (27.5–50%) cell proliferation and reduced cell migration in all strains evaluated. Additionally, the lectin decreased the invasion of SCC9 cells. Apoptosis was higher against B16 and SCC9 cells treated with the lectin. GaBL induced the upregulation of caspase-3, E-cadherin and suppression of type I collagen in all strains tested, indicating lower cancer development. GaBL induces inhibition of growth, migration, invasion, and regulation of the expression of caspase-mediated apoptosis and proteins related to the development of cancer in human head and neck cells.