The three-fold objectives of the current study were to: (i) investigate both the antioxidant and in vitro anticancer activities of chemically characterized ethanolic extract of Centaurea calcitrapa (EECC) aerial parts (leaves and roots), (ii) elucidate the EECC-mediated molecular mechanism underlying its cytotoxicity against breast cancer cells by use of FACS and (iii) conduct in silico analysis of its lead compounds for the assessment of their inhibitory activity against caspase-3, the executioner enzyme of apoptosis, by use of molecular docking. This evidence concerns the gene CASP3 and breast carcinoma.