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Background Penfluridol, a commonly used antipsychotic agent in a clinical setting, exhibits potential anti-cancer properties against various human malignancies. Here, we investigated the effect of penfluridol on the biological behavior of CRC cells. Methods Cell viability and clonogenic potential were detected by the cell counting kit-8 and colony formation assay. The cell apoptosis and cell cycle distribution were quantified through flow cytometry. Caspase-3 activity, glucose consumption, lactate production, and intracellular ATP levels