Colorectal cancer (CRC) is one of the most prevalent malignancies. Accumulating evidence suggests that Hippo signaling pathway is a crucial signaling pathway that regulates stemness in multiple cancers, including CRC. Given that, the therapeutics targeting the Hippo pathway are of great interest. At the same time, previous studies have reported that PP2A regulates the Hippo pathway in tumorigenesis. In this study, we uncover the role of PP2A in vitro and in vivo and the interaction between PP2A and Hippo signaling pathway in CRC stemness. We investigate the molecular mechanism of PP2A-mediated Hippo pathway in CRC progression from the aspect of cell stemness. Furthermore, we explore the transcription factor of PP2A in CRC. In view of circRNA's regulation of cancer-related signaling pathways in CRC, we also study the association between PP2A-mediated Hippo pathway and the CRC-related circRNA (hsa-circ-001680) confirmed in previous studies. Of note, we demonstrate how hsa-circ-001680 regulates PP2A expression. The effect of hsa-circ-001680 on PP2A stability is investigated as well. In conclusion, the study demonstrates the mechanism of PP2A-mediated pathway on CRC stemness, which may provide new sights for the treatment of CRC.