Network Pharmacology, Molecular Docking, and Wnt Signaling-Associated Gene Expression Modulation Effects of New...

Colorectal Cancer (CRC) remains a major global health challenge due to the emergence of resistance to current therapies. Identifying selective compounds that effectively trigger apoptosis in cancer cells while sparing normal tissues is therefore essential.

Candidate molecules were identified through virtual screening and network-based prioritization targeting p53, BAX, and BCL-2. Molecular docking and Molecular Dynamics (MD) simulations were used to validate binding stability and target specificity. In vitro assays were conducted on human CRC (HCT116) and normal colon fibroblast (CCD-18Co) cells to evaluate cytotoxicity, proliferation, and DNA fragmentation. QRTPCR performed gene expression analyses of p53, BAX, and BCL-2 following treatment with 25-100 μg/mL of each compound for 12-72 h.

Among the screened compounds, LC_60 emerged as the most potent and selective candidate, exhibiting a remarkably low IC₂⁽ (0.0128 μg/mL) toward HCT116 cells and negligible toxicity toward CCD-18Co cells (CC₂⁽ = 125 μg/mL), yielding an exceptionally high selectivity index (SI ≈ 9,766). Docking and MD simulations confirmed strong and stable interactions of LC_60 with BAX and β-catenin, supporting its dual role in apoptosis induction and Wnt pathway inhibition. LC_60 treatment significantly upregulated p53 and BAX while downregulating BCL-2, consistent with activation of intrinsic apoptotic signaling.

LC_60's pronounced potency and outstanding selectivity underscore its ability to selectively modulate apoptosis in malignant colon cells while sparing normal fibroblasts, suggesting a strong therapeutic window and tumor specificity.

Collectively, the findings identify LC_60 as a highly selective, multitargeted lead compound that activates p53-BAX-mediated apoptosis and inhibits oncogenic Wnt signaling, establishing it as a promising scaffold for further preclinical development in colorectal cancer management.