Dysregulation of the Hippo pathway has been frequently identified in various human cancers. The Hippo pathway is a highly complex pathway involving multiple types of proteins, and the activation of YAP by LATS kinase is the final effector step in the transcription process. In this study, we linked the roles of the multifunctional adapter proteins β-arrestins (barrestin 1 and 2) in cooperation with other signaling pathways such as GPCR and Wnt to essential cellular functions involved in carcinogenesis, including the regulation of cell proliferation, migration, and differentiation as well as stem cell properties. Although β-arrestin 1 and 2 have high structural similarities, they have different roles in carcinogenesis as β-arrestin 1 aids cancer cell survival and metastasis and β-arrestin 2 inhibits tumor growth. In the Hippo signaling pathway, β-arrestins function as scaffold proteins that mediate the phosphorylation of key molecules, and their association with human cancers is a major research topic. In this study, we demonstrated that β-arrestin 2 inhibits YAP activation through the formation of the β-arrestin 2-LATS-YAP trimeric complex, which results in the promotion of the kinase activity of LATS in cancer cell lines and patient-derived colon cancer organoids.