Background: Breast cancer (BC) is the most diagnosed cancer among women worldwide. Despite available chemotherapeutic drugs effective against cancer, BC remains a complex disease with high mortality incidence. Scorpion venom (SV) is a poisonous mixture that contains thousands of bioactive proteins that previously demonstrated pharmacological properties, including anticancer potentials. Our present study investigated the antitumor effect of the venom of Egyptian scorpions: Androctonus australis and Androctonus bicolor from Marsa Matrouh. The IC50 dose of venom was determined through MTT assay, followed by an evaluation of the effect of venoms on cancer cell progression, cell cycle arrest, apoptosis, and DNA fragmentation. In-vitro cytotoxicity of venom was evaluated against MDA-MB-231 and MCF-7 cancer cells, with HSF normal cells. SV showed significant inhibition of cancer cell viability (32__52.56 % inhibition at the highest tested concentration of 200μg/mL), respectively. Aa and Ab venoms were cytotoxic to cancer cells, where Aa venom with a prominent inhibitory effect on MCF-7, while Ab venom with a prominent inhibitory effect on MDA-MB-231 cells. No signs of toxicity were observed on HSF cells. Venoms significantly prevented cancer cell progression while exhibiting insignificant effects on HSF cells. The apoptotic pathway was evaluated using Annexin-V/FITC & PI staining and agarose gel. Venoms caused a DNA damage-mediated cell death in cancer cells then confirmed using RT-qPCR that showed upregulation of P53, P21 tumor suppressor gene, and Bax apoptotic gene, and downregulation of Bcl-2 anti-apoptotic gene. SDS-PAGE results revealed the presence of various peptides that ranged from 25kDa to 60kDa. Peptide bands of 245kDa, 90kDa, 75kDa, and 40kDa were found common in venom 1 and venom 2, while peptide bands of 100kDa, 48kDa to 60kDa, 25kDA, 17kDa, and 11kDa were shared between venom 1 and venom 2. One unique peptide band suggests inter-genus and inter-species relationships among different species and explains their various therapeutical potentials. Our results suggest that scorpion venom exhibits selective cytotoxic properties via apoptosis induction in breast cancer.