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Objective: To investigate the cytotoxic, antioxidant and antibacterial activities of ethanolic and aqueous crude extracts of Nerium oleander (N. oleander) leaves. Methods: Cytotoxic activities were evaluated by WST-1 bioassay on two human cancer cell lines, namely human colon adenocarcinoma cell line HT-29 and human breast cancer cell line MDA-MB-231. The antioxidant property of N. oleander extracts was assessed by DPPH scavenging and β-carotene bleaching tests. The agar disc diffusion method was used for the determination of antimicrobial activity against different strains. Results: Using cell viability indices, the WST-1 test revealed that both extracts reduced cell viability in both cell lines. Our results also showed that aqueous extract was more active than ethanolic extract, with IC50 values of (1.67±0.22) μg/mL and (2.36±0.44) μg/mL on MDA-MB-231 cells, and (2.89±0.35) μg/mL and (5.09±0.52) μg/mL on HT29 cells, respectively. The study of the antioxidant activity showed that N. oleander extracts had a considerable scavenging capacity and exerted a significant preventive effect against the oxidation of β-carotene by the peroxide radicals. However, the antibacterial test showed thatboth ethanolic and aqueous extracts of N. oleander had a moderate antibacterial effect limited only to Gram-positive bacteria. Conclusions: Our results shows that N. oleander aqueous and ethanolic extracts have significant cytotoxic activities against tumor cell lines and possesses a strong antioxidant capacity, suggesting the presence of active compounds in N. oleander leaves that could be a potential source of phytochemicals with high medicinal value to be used in cancer treatment and prevention.