Colorectal cancer (CRC) is a leading cause of cancer mortality, primarily due to metastasis and treatment refractoriness. These challenges have prompted the search for novel natural compounds, including 20(S)-protopanaxadiol (PPD), a key metabolite of Panax ginseng ginsenosides. While PPD has shown potent anti-tumor activity, its efficacy against both chemosensitive and chemoresistant CRC proliferation and dissemination remains unclear. To address this gap, our study investigates the anti-proliferative and anti-metastatic potential of PPD using murine CRC cell lines: the wild type CT26 cells and a derivative line with acquired resistance to 5-fluorouracil (5-FU). PPD potently suppressed the proliferation and clonogenic survival of both chemosensitive and 5-FU-resistant CRC cells. This cytostatic effect was associated with G0/G1 phase cell cycle arrest, mediated by the downregulation of cyclin D1 and CDK4. In addition, PPD activated multiple cell death pathways, including autophagy, marked by increased LC3B and p62 levels, and apoptosis, which was initiated through mitochondrial depolarization. In a murine model of metastasis, oral administration of PPD effectively reduced the formation of pulmonary tumor nodules. Taken together, our data suggest that PPD impedes the progression of CRC by restricting cell growth while activating apoptosis and autophagy. These results highlight PPD as a promising therapeutic candidate for the treatment of metastatic CRC.