Abstract
Bisabolol oxide A, a key bioactive constituent of Matricaria chamomilla, is recognized for its potent anti-inflammatory, antioxidant, and anticancer properties. This study focused on synthesizing bisabolol oxide A-loaded micellar/liposomal nanoparticles (NPs) and rigorously evaluating their anti-proliferative and anti-invasive effects using integrated in vitro and in silico methodologies. Physicochemical characterization of the bisabolol oxide A-loaded NPs was performed using SEM, TEM, TGA-DTG, Zeta potential, DLS analysis, and FT-IR spectroscopy. Furthermore, drug-like properties for bisabolol oxide A, bisabolol oxide B, and 5,8,11-heptadecatriynoic acid methyl ester were predicted using Swiss ADME software. Molecular dynamics simulations were subsequently conducted for these compounds against critical cellular migration proteins: β-catenin and the Epidermal Growth Factor Receptor (EGFR). The anticancer efficacy of the bisabolol oxide A-loaded nanoparticles was assessed through MTT assays, Annexin V/PI staining, wound healing assays, and qRT-PCR. Characterizations confirmed that the nanoparticles possessed optimal characteristics, exhibiting sizes between 35 to 67 nm, high mono-dispersity and thermal stability up to 380°C. In vitro results demonstrated that bisabolol oxide A-loaded micellar/liposomal nanoparticles significantly inhibited DU-145 cell proliferation, yielding IC₅₀ value of 55.44 µg/mL, 46.21 µg/mL, and 43.59 µg/mL after 24, 48, and 72 hours of treatment. Flow-cytometry analysis further confirmed 50.04% reduced migratory capacity and induction of apoptosis. Mechanistically, qRT-PCR analysis revealed a significant 2.31 ± 0.16-fold upregulation of the tumor-suppressive miR-34a and a 1.72 ± 0.03-fold downregulation of the oncomiR miR-181a in treated cells. These miRNA alterations correlated with the downregulation of migration-associated genes (CTNNB, SMAD3, and EGFR) and the upregulation of apoptosis-related genes (P53 and CASP9), findings substantiated by Western blot analysis. In silico analyses indicated favorable ADMET properties for bisabolol oxide A, bisabolol oxide B, and 5,8,11-heptadecatriyneoic acid methyl ester. Molecular docking simulations revealed strong binding affinities of these compounds for β-catenin and EGFR, which are key proteins in cellular migration pathways. Our findings strongly suggest that bisabolol oxide A functions as an anti-migration and pro-apoptotic agent by effectively modulating the expression of specific miRNAs and their transcriptional targets, complemented by potent inhibitory interactions with critical protein targets.