Validation of herbal flavonoids for breast cancer through pharmacological networking and <i>in-vitro</i> studies against MCF-7 and HepG2

Breast cancer, the most common malignancy in developed countries, involves 12% of 20–34-year-old females. Herbal treatments for breast cancer are growing popular due to adverse effects of conventional treatments. Network pharmacology, ethnobotany, and in vitro studies have described bioactive compounds found in plants grown in Pakistan’s Southern Punjab for treating breast cancer. A quantitative analysis for selection of five plants was performed. Plant samples were collected, dried, prepared, and mounted on herbarium sheets. Phytochemicals were evaluated for chemical absorption, distribution, metabolism, excretion, and toxicity properties and target using SwissTargetPrediction. Potential breast cancer targets were identified via GeneCards database and database of gene disease association (DisGeNET). Enrichment analysis and protein interactions were exercised via database for annotation, visualization, and integrated discovery (DAVID) and Cytoscape. A drug compounds–genes–disease network found key genes for treatment, validated by molecular docking. The top three docking complexes withstood 200-nanoseconds (ns) molecular dynamics simulations in GROningen MAchine for Chemical Simulations (GROMACS) 2020, showing average Coulombic short-range interaction energies of apigenin with androgen receptor (-30.64 kJ/mol), apigenin with estrogen receptor 1 (ESR1; -62.35 kJ/mol), and luteolin with epidermal growth factor receptor (EGFR; -95.99 kJ/mol). Cytotoxicity of five plant extracts was analyzed on HepG2 and MCF7 cell lines. Liquid chromatography–mass spectrometry was performed for the compounds that had best half-maximal inhibitory concentration (IC50) value. This research investigates the anti-breast cancer mechanisms of plant flavonoids at molecular level.