Objective In this study, we analyzed and investigated the antioxidant, anti-inflammatory, and antitumoral properties of methanolic (Met), dichloromethane (Dic), ethyl acetate (Ac), and diethyl ether (Et) fruit extracts of Ammodaucus leucotrichus on the MCF-7 breast cancer cell line. Methods The extracts of Ammodaucus leucotrichus fruits, explored in this study, were obtained by cold maceration and analyzed by LC-MS. 1,1-diphenyl-2-picrylhydrazyl (DPPH), β-Carotene and Bovine Serum Albumin (BSA) assays were used to assess antioxidant and anti-inflammatory activities. The antiproliferative and proapoptotic effects of extracts were determined by the Resazurin test and quantitative real-time polymerase chain reaction (qRT-PCR) analysis. Results Phytochemical screening of Ammodaucus leucotrichus fruit extracts revealed the presence of flavonoids (luteolin and cynaroside), iridoid derivatives, phenolic acids, and identified Rhodojaponin—a major cytoxic compound. The DPPH and BSA assays revealed the most potent radical-scavenging activity (IC 50 = 0.16 mg/mL) and anti-inflammatory effect (IC 50 = 35.87 μg/mL) in Met extract. Met, Dic, Ac, and Et extracts, at 100 µg/mL, exhibited significant antiproliferative activity against MCF-7 reaching inhibition percentage of 34%, 31%, 28%, and 13%, respectively. qRT-PCR analysis was marked by the inhibition of proliferative and inflammatory markers Proliferating Cell Nuclear Antigen (PCNA) and IL-6 and the up-regulation of antioxidant and proapoptotic genes Nrf2, Bax, and Caspase 3 in Met and Dic extracts, without significant changes to anti-apoptotic gene Bcl-2 expression. Conclusion The study revealed the potential therapeutic effects of Ammodaucus leucotrichus fruit extracts, suggesting they could be used as an adjuvant to breast cancer therapies.