Cancer is one of the main causes of mortality and morbidity worldwide, which does not have a fully effective medical treatment. Therefore, the objective of this study was to evaluate the effect of lipidically characterized and hydrolyzed chia seed (Salvia hispanica L.) oil on in vitro colon cancer (Caco2 cell line) and breast cancer cells (MCF7 cell line) cell viability. Cancer cells were treated with different hydrolyzed oil concentrations (12.5-400 mu g/mL) for 48 h; then cell viability was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay. All chia seed oil concentrations significantly reduced cell viability in the Caco2 line, with the best reduction being 23.88% with the 25 mu g/mL concentration. However, in the breast cancer line the highest concentrations (200 and 400 mu g/mL) increased the viability of malignant cells. Chia seed oil significantly reduces the viability of Caco2 cells, although at the higher concentrations it can increase the viability of breast cancer cells.