Retinol utilizes a retinoid X receptor (RXR)-mediated degradation pathway to decrease β-catenin protein in all-trans retinoic acid (ATRA)-resistant human colon cancer cells. In this study, we examined interactions between RXRα and β-catenin in ATRA-resistant human colon cancer cells treated with retinol. Retinol treatment triggers relocation of β-catenin and RXRα proteins. Cells treated with retinol for 8 and 24 h displayed increased cytosolic but decreased nuclear β-catenin and RXRα. Retinol treatment increased β-catenin and RXRα protein interaction. Previously, we showed that 24 h of retinol treatment increased RXRα protein. Here we show this increase in RXRα levels is due to increased RXRα messenger RNA. Treatment with 48 h with retinol decreased RXRα protein levels. Last, by transfecting HCT-116 cells with a RXRα construct lacking the activation function-1 and DNA binding domains, we show RXRα and β-catenin binding is required for proteosomal degradation of β-catenin. These results suggest retinol induces RXRα and β-catenin binding and transport to the cytosol where they are proteasomally degraded.