Khang Wen Goh0000-0001-5422-5123Jie JiXian GuLokesh Bontha Venkata SubrahmanyaHemant Kumar Singh Yadav Singh YadavPhei Er Kee0000-0001-9299-6842Siew-Keah LEEAshok Kumar JanakiramanLong Chiau MingKai Bin Liew2026-10-212026-10-212025-09-1810.36877/pmmb.a0000472https://dspace-cris.utar.edu.my/handle/123456789/11809Colorectal cancer (CRC) remains a major global health burden, necessitating alternative therapeutic approaches. Traditional Chinese Medicine (TCM), including BuShenFang (BSF), has demonstrated potential anti-CRC effects, yet its precise mechanisms remain unclear. While BSF has previously exhibited anti-CRC activity in vitro, its in vivo efficacy and associated molecular mechanisms requires further elucidation. In this study, a subcutaneous xenograft model was established in nude mice, which lack T-cell-mediated immunity and can effectively support the growth of human colorectal cancer cells (HCT116). This model enables a more clinically relevant evaluation of BSF’s antitumor effects in vivo. The study aimed to assess the tumor inhibitory effect of BSF and investigate its underlying mechanisms via the Wnt/?-catenin signaling pathway. Mice were allocated into control, low-dose BSF (L-BSF), high-dose BSF (H-BSF), and xStAxVHLL (positive control) groups. Tumor growth was monitored, and analyses including histology, immunohistochemistry, TUNEL assay, immunofluorescence, and Western blotting were performed to examine tumor proliferation, apoptosis, and Wnt pathway regulation. BSF significantly inhibited tumor growth in a dose-dependent manner, reduced KI67 expression, promoted apoptosis, and increased adenomatous polyposis coli (APC) levels. Additionally, BSF downregulated ?-catenin, p-GSK-3?/GSK-3?, and Axin1, while suppressing Wnt target genes (Cyclin D1, c-Myc, COX-2). The decrease in p-GSK-3? (Ser9) suggests increased GSK-3? activity, which promotes ?-catenin degradation and attenuates Wnt/?-catenin signaling. These indicate that BSF inhibits CRC progression through modulation of the Wnt/?-catenin signaling pathway, promoting apoptosis and suppressing tumor proliferation. This highlights its potential as an adjunct therapy in CRC treatment. In conclusion, these findings suggest that BSF may exert anti-CRC effects by targeting key tumorigenic pathways in preclinical study. Further research, including clinical trials, is needed to confirm its therapeutic potential. © 2025, HH Publisher. All rights reserved.en-USBuShenFangColorectal cancerNude miceSDG 3 Good health and well-beingSubcutaneous Transplanted TumorTraditional Chinese Medicinejournal-article