This study aimed to investigate the potential anticancer properties of Cannabis sativa extracts, particularly focusing on the identification and characterization of individual compounds. Additionally, the research aimed to assess the molecular impact of quercetin on colorectal cancer cells. Cannabis sativa leaves were collected, and extracts were obtained using different solvents (hexane, chloroform, methanol, and water). Phytochemical analysis, including the detection of alkaloids, flavonoids, phenols, glycosides, tannins, carbohydrates, saponins, and steroids, was conducted. The total flavonoid content was determined using the aluminum chloride colorimetric method. Antioxidant activity was evaluated through DPPH, FRAP, and H2O2 assays. The extracts exhibiting anticancer activity were further purified through column chromatography and HPLC. The isolated compounds were tested for their anticancer potential against HCT116 colorectal cancer cells, and IC50 values were calculated. Morphological analysis and gene expression studies were performed to assess the cellular and molecular impact of the identified compounds. The water extract showed the highest extraction yield and a rich phytochemical profile. Methanol and water extracts displayed robust antioxidant activity. Column purification and HPLC led to the isolation of quercetin, which exhibited significant anticancer activity against HCT116 cells with an IC50 of 60.253 µg/ml. Morphological analysis revealed dose-dependent cytotoxicity. Gene expression analysis indicated an upregulation of M-CSF and a slight upregulation of GM-CSF in quercetin-treated cells. This study provides insights into the diverse chemical compositions and biological activities of Cannabis sativa extracts. The identified quercetin compound demonstrated promising anticancer properties against colorectal cancer cells, warranting further investigation into its therapeutic potential. The molecular analysis sheds light on the potential mechanisms underlying quercetin's effects, contributing to the understanding of its role in cancer treatment