Skin cancer is one of the most aggressive types of neoplasms, with high morbidity and mortality. Numerous factors are associated with the development of melanoma, including exposure to UV radiation and the use of exogenous hormones. Some studies suggest that oral contraceptives can influence the development of skin cancer, but the evidence is insufficient and contradictory. Our study aimed to initially evaluate the safety profile of the combination of levonorgestrel and 17β-ethinylestradiol on healthy cell lines (HaCaT and JB6 Cl 41-5a) and to further determine the possible association with skin cancer by investigating the effect of the combination on the murine melanoma tumor line (B164A5). The data obtained showed that levonorgestrel-ethinylestradiol (LG-EE) did not have a toxic effect on healthy cells (HaCaT and JB6 Cl 41-5a) but exhibited a slight proliferative effect on murine skin tumor cells at increasing concentrations. LG-EE on healthy lines did not significantly decrease viability and did not induce a cytotoxic effect. On the B164A5 tumor line, the hormonal combination at a concentration of 5 µM slightly decreased viability and degraded the cell membrane, observing a decrease in confluence and number of cells, as well as the presence of apoptotic bodies, while at the highest dose tested of 15 µM, an opposite effect was recorded with a slight stimulation of murine tumor cells. The results suggest that LG-EE may influence the development of melanoma; however, the evidence is insufficient, and further studies are necessary.