Objective To explore the impact of proguanil and osimertinib, both as individual treatments and in combination, on the migratory behavior of bladder cancer cells, as well as to elucidate the potential mechanisms underlying these effects.Methods MTT assays were conducted to assess the impact of proguanil and osimertinib as single agents on the proliferation of bladder cancer cells. Cell scratch assays and Transwell assays were employed to evaluate the effects of proguanil and osimertinib, both individually and in combination, on the migration of bladder cancer cells. Western blotting analysis was used to examine the effects of proguanil and osimertinib in combination on the expressions of epidermal growth factor receptor (EGFR) and epithelial-mesenchymal transition (EMT) markers, specifically E-cadherin and N-cadherin.Results Monotherapy with either proguanil or osimertinib significantly suppressed the proliferation and migration of bladder cancer cells. The combination treatment of proguanil and osimertinib demonstrated a marked synergistic inhibiting effect, resulting in enhanced inhibition of both cell proliferation and migration compared to single-agent treatment. The combination treatment significantly inhibited EGFR phosphorylation, downregulated N-cadherin expression, and upregulated E-cadherin expression.Conclusion The combination of proguanil and osimertinib effectively suppresses the proliferation and migration of bladder cancer cells via inhibiting EGFR phosphorylation and blocking the EMT process.