Colon cancer is a common malignancy of the digestive tract. Colon cancer stem-like cells (CCSCs) are theoretically one of the key drivers of the initiation, relapse, metastasis, and chemo-resistance of colon tumors. Piezo1 is a mechanosensitive cationic channel protein involved in cancer progression. However, little is known regarding the possible role of Piezo1 in maintaining the stemness of CCSCs. In this study, we found that Piezo1 was highly expressed in CD133+/CD44+colon cancer tissues, and the Piezo1high/CD133+CD44+population was associated with the clinical stage. Furthermore, CCSCs isolated from colon cell lines expressed higher Piezo1 levels compared to the non-CCSCs, and Piezo1 knockdown inhibited their tumorigenicity and self-renewal capacity. Mechanistically, Piezo1 maintained the stemness of CCSCs through Ca2+/NFAT1 signaling, and knocking down Piezo1 promoted degradation of NFAT1. Taken together, Piezo1 is involved in the stage of colon cancer and is a promising therapeutic target.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12672-023-00712-4.