The present study established thirteen novel 8-hydroxyquinoline/chalcone hybrids 3a–m of hopeful anticancer activity. According to NCI screening and MTT assay results, compounds 3d- 3f, 3i, 3k, and 3l displayed potent growth inhibition on HCT116 and MCF7 cells compared to Staurosporine. Among these compounds, 3e and 3f showed outstanding superior activity against HCT116 and MCF7 cells and better safety toward normal WI-38 cells than Staurosporine. The enzymatic assay revealed that 3e, 3d, and 3i had good tubulin polymerization inhibition (IC50 = 5.3, 8.6, and 8.05 µM, respectively) compared to the reference Combretastatin A4 (IC50 = 2.15 µM). Moreover, 3e, 3l, and 3f exhibited EGFR inhibition (IC50 = 0.097, 0.154, and 0.334 µM, respectively) compared to Erlotinib (IC50 = 0.056 µM). Compounds 3e and 3f were investigated for their effects on the cell cycle, apoptosis induction, and wnt1/β-catenin gene suppression. The apoptosis markers Bax, Bcl2, Casp3, Casp9, PARP1, and β-actin were detected by Western blot. In-silico molecular docking, physicochemical, and pharmacokinetic studies were implemented for the validation of dual mechanisms and other bioavailability standards. Hence, Compounds 3e and 3f are promising antiproliferative leads with tubulin polymerization and EGFR kinase inhibition.