Prostate cancer (PCa) is the most prevalent cancer in men who live in developed nations. Even with the recent advancements in surgery, radiation, hormonal therapy, and medical management, medical treatment for advanced-stage PCa is being developed. Alternative strategies for reducing the prevalence of PCa include dietary cancer chemopreventive agents, it has been suggested that medicinal plants can treat cancer. Active composites of Capparis Cartilaginea (C. cartilaginea) are alkaloids, tannins, steroids, terpenoids, and flavonoids. This study aims to investigate the cytotoxic effect of C. cartilaginea fruit extract on the 22RV1 human prostate cancer cell line. Under an inverted light microscope, a dose- and time-dependent morphological alteration and cell migratory capacity were observed after exposure to the ethanol extract of C. cartilaginea fruit. The cytotoxicity of the C. cartilaginea fruit ethanol extract on the prostate cancer 22RV1 cells was performed by using the 3-4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide colorimetric assay. The effect of C. cartilaginea fruit extract on cell migration was assessed using an in vitro wound-healing scratch assay. The MTT assay showed that C. cartilaginea fruit extract had a significant cytotoxic effect on 22RV1 cells after 72 h treatment with a value of (0.088 ± 0.031) μg/mL. Following exposure to C. cartilaginea fruit extract, morphological changes in the cells showed that the extract had a remarkable impact on cell death, and the mortality correspondingly increased with the increase in concentration. The findings suggested that C. cartilaginea fruit ethanol extract might be used as a medicinal plant for treating prostate cancer.