Abstract 6330: Unveiling the anticancer potential of <i>Rheum webbianum</i> in colorectal cancer defense through modulation of key wnt/β-catenin and TGF-β signaling pathways: An in-vitro and in-vivo investigation

Abstract Purpose of the Study: Colorectal cancer (CRC) remains a significant health concern worldwide. Lifestyle and environmental factors are primary contributors to CRC, emphasizing the need for effective preventive strategies. Rheum webbianum (RW), a plant with a rich ethnopharmacological background, has demonstrated promising chemopreventive potential. In our previously published study on the ameliorative potential of Rheum webbianum extracts in DMH-induced colorectal cancer (U.M. Khaja et al., 2024), the ethanolic extract of RW demonstrated the highest ameliorating potential in both in-vitro and in-vivo experiments. Building on these findings, this study aims to further investigate the anticancer effects of RW ethanolic extracts through additional in-vitro and in-vivo assays, focusing on their impact on critical molecular pathways involved in CRC progression. Materials and Methods: The study evaluated the anticancer potential of RW ethanolic extracts using the HT-29 human colon cancer cell line and a 1,2-dimethylhydrazine (DMH)-induced rat CRC model. In vitro assays assessed the effects of RW on cancer cell viability and migration, using cytotoxicity and Transwell migration assays. Western blot analysis was conducted to explore the extract's influence on key signaling pathways, including Wnt/β-catenin and TGF-β, both of which are critical drivers of CRC progression and metastasis. Results: In-vitro: The ethanolic extract of Rheum webbianum (RW) significantly inhibited the viability of HT-29 cells in a dose- and time-dependent manner, with up to 80% inhibition observed at a concentration of 200 µg/mL. Transwell migration assays demonstrated a substantial reduction in cancer cell migration, with progressively stronger inhibitory effects seen at higher RW concentrations. In-vivo: In the DMH-induced rat CRC model, molecular analysis revealed that RW extracts suppressed β-catenin expression, indicating inhibition of the Wnt/β-catenin signaling pathway, a hallmark of CRC progression. Furthermore, the extracts downregulated TGF-β1 expression, implicating their role in reducing pro-metastatic signaling. These findings highlight the extract’s ability to target critical pathways involved in tumor growth and metastasis. Conclusion: This study provides compelling evidence of the anticancer potential of RW ethanolic extracts. The results highlight their ability to inhibit cell viability, suppress migration, reduce metastatic behavior, and modulate key signaling pathways associated with CRC progression. These findings emphasize the potential of RW as a natural, plant-based chemopreventive agent and pave the way for future research to isolate active bioactive compounds and explore their applications in cancer prevention and integrative treatment strategies. Citation Format: Umer Majeed Khaja, Reena Singh, Showkat Ahmad Ganie. Unveiling the anticancer potential of Rheum webbianum in colorectal cancer defense through modulation of key wnt/β-catenin and TGF-β signaling pathways: An in-vitro and in-vivo investigation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 6330.