Abstract
Worldwide, colorectal cancer continues to be a major contributor to cancer mortality. The main reasons for high-colorectal cancer death rates are tumor progression, metastasis, and resistance to existing therapies. Wnt/β-catenin and PI3K/AKT/mTOR pathways have been shown to be involved in the development of colorectal cancer as well as in resistance to available therapies. Therefore, agents that can target both pathways simultaneously would be advantageous. Baicalein, a bioactive flavonoid from the plant Scutellaria baicalensis, has been shown to have anticancer activity against multiple cancers, but its dual effects on the Wnt/β-catenin and PI3K/AKT/mTOR pathways in human colorectal cancer remain undefined. In this study, we aimed to determine whether baicalein has antitumor activity and the molecular mechanism through which it acts in human colorectal cancer cell lines SW480 and HCT116. We assessed baicalein's antitumor activity using PrestoBlue cell viability, flow cytometry for determining apoptosis, wound-healing assay for migration, and colony formation assay for determining clonogenic survival. The molecular mechanisms by which baicalein inhibits tumors were evaluated using various molecular techniques including reverse transcription-quantitative polymerase chain reaction (RT-qPCR), Western blotting, and nuclear-cytoplasmic fractionation to measure the activity of the Wnt/β-catenin and PI3K/AKT/mTOR pathways. In addition, the clinical relevance of baicalein's targets was evaluated using in silico analysis of the Cancer Genome Atlas (TCGA) dataset. The compound baicalein decreased proliferation, migration and colony forming ability of colorectal cancer cells and increased apoptosis. These actions were dependent upon time and concentration of baicalein treatment. The anti-proliferative actions of baicalein occurred through inhibition of Wnt/β-catenin signaling by decreasing AXIN2 and GSK3β expression and nuclear accumulation of β-catenin. Baicalein also inhibited PI3K/AKT/mTOR signaling by decreasing mTOR expression as well as phosphorylation of AKT, mTOR, and S6. In addition, the results of the in silico analyses suggested that these targets may be clinically relevant and correlate with drug resistance. Therefore, baicalein is identified to be a potent dual inhibitor of Wnt/β-catenin and PI3K/AKT/mTOR signaling pathways and provides evidence for its use as a novel therapeutic agent for colorectal cancer. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10616-026-00959-z.