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The application of various agricultural pesticides can lead to adverse effects on bodily tissues. This study evaluated the impact of dermal exposure to malathion on sex hormone levels and its potential carcinogenicity in mice. 30 adult male mice were randomly assigned to control and treatment groups. The treatment group received a topical application of malathion at 1 ppm (1 ml) for 12 weeks. After the exposure period, serum levels of testosterone, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) were analyzed, and tissue samples were collected post-necropsy. Cytotoxicity was assessed using the micronucleus assay and MTT method in vitro, and the IC50 value was calculated. Results showed no significant difference in LH levels between groups; however, both testosterone and FSH levels declined significantly in the treated mice. Histological evaluation revealed mild liver toxicity in the treatment group. Additionally, the frequency of micronuclei at various Malathion concentrations was significantly elevated compared to controls (P < 0.05). Overall, these findings suggest that dermal exposure to malathion can reduce reproductive hormone levels and induce cytotoxicity in mice.