Integrated computational and in vitro investigation to explore the preliminary anti-proliferative and anti-mitotic potential of γ-oryzanol against breast cancer.

This study investigated the preliminary role of γ-oryzanol (γ-O) against breast cancer through computational and in vitro analyses. Reference protein set (RPS) and functional enrichment set (FES) proteins were identified and subjected to network pharmacology. Pharmacogenomics analysis assessed expression profiles and survival associations. Molecular docking of γ-O with PIK3CA, AKT1, AKR1C3, ESR1, and MAPK1 was performed, followed by molecular dynamics (MD) simulation of the γ-O-PIK3CA complex. Brine shrimp lethality assay (BSLA) evaluated the effect of γ-O on nauplii survival and percentage lethality. Allium cepa root tip assay (ACARTA) assessed the effect of γ-O in number and length of roots. Cell viability assay evaluated MCF-7 cell viability after γ-O treatment. Target validation and network pharmacology suggested the relevance of RPS and FES with cancer-associated pathways. Pharmacogenomics analysis highlighted FES overexpression with reduced survival associations and cancer progression. Molecular docking predicted interaction of γ-O with PIK3CA (- 9.1 kcal/mol), AKT1 (- 7.2 kcal/mol), AKR1C3 (- 12.3 kcal/mol), ESR1 (- 8.3 kcal/mol), and MAPK1 (- 8.9 kcal/mol). MD simulation indicated the stability of γ-O-PIK3CA complex. γ-O treatment significantly (P-value < 0.05) reduced nauplii survival and increased percentage lethality. The number and length of roots reduced significantly (P-value < 0.05) in γ-O-treated roots. γ-O produced a significant (P-value < 0.05) concentration-dependent reduction in cell viability (half-maximal inhibitory concentration = 352.57 ± 48.51 µg/mL). BSLA, ACRTA, and cell viability assay demonstrated preliminary cytotoxic, anti-mitotic, and anti-proliferative effects of γ-O, aligned with computational target and interaction analyses. Further studies are required to validate its mechanistic relevance against breast cancer.