Abstract
Echinops L. is recognized for its anticancer and immunomodulatory properties. This study is the first to investigate the potential anticancer and immunomodulatory effects of Echinops shakrokii S.A. Ahmad, collected from Kurdistan, Iraq, employing both in vitro and in vivo methods. Solvent extracts (ethyl acetate, aqueous methanol, and aqueous) from the aerial parts of the plant were prepared via maceration and Soxhlet extraction. The MTT assay assessed antiproliferative efficacy against the MCF-7, MDA-MB-231, EMT6/P, HeLa, T47-D, Caco-2, and VERO cell lines, with results compared to doxorubicin as a positive control. Apoptosis induction was evaluated using an ELISA kit. Immune system activation was assessed through assays of lymphocyte proliferation and macrophages' pinocytic and phagocytic activities. LC-MS identified phytochemicals in the three extracts. EMT6/P cancer cells were injected intraperitoneally into Balb/c mice to investigate the anticancer activity of the aqueous methanol extract. Liver and kidney toxicities were also evaluated. Due to the presence of various phytochemical components, including flavonoids, phenolic compounds, alkaloids, and terpenoids, the ethyl acetate and aqueous methanol extracts exhibited significant antiproliferative effects. The aqueous methanol extract demonstrated the highest potency against the T47D cell line (IC(50) = 0.50 ± 0.019 mg/ml), while the ethyl acetate extract displayed the maximum activity against the MCF-7 cell line (IC(50) = 0.56 ± 0.03 mg/ml). In vivo, the aqueous methanol extract healed 40% of the mice and reduced the average tumor size by 39.29%. There was no significant increase in renal or hepatic toxicity markers at a dose of 675 mg/kg/day, which was selected based on the acute toxicity test. The plant induced apoptosis through caspase-3 activation. Lymphocyte proliferation, macrophage phagocytosis, and pinocytosis were also enhanced. These findings position E. shakrokii as a potential natural anticancer agent with dual apoptotic and immunostimulatory properties. Its mechanisms should be further investigated for their possible use in cancer therapy regimens.