Inhibitory Effects of Hesperetin and EGCG in the Lung Cancer Progression involves impairment in TOP2A gene expression regulation.

Lung cancer (LC) remains the leading cause of death among all cancer-related mortalities. Despite advances in Non-Small Cell Lung Cancer (NSCLC) treatment, rising drug resistance necessitates the development of new therapeutics targeting relevant biomarkers. Understanding resistance mechanisms is key to improving treatment effectiveness and overcoming drug resistance. This study aims to identify crucial biomarkers and evaluate potential drug-like compounds targeting them using both in silico and in vitro analyses. Ten key biomarkers associated with LC-including Assembly Factor for Spindle Microtubules (ASPM), Budding Uninhibited by Benzimidazoles 1 (BUB1), Centromere Protein F (CENPF), DNA Topoisomerase II Alpha (TOP2A), TPX2 Microtubule Nucleation Factor (TPX2), Kinesin Family Member 15 (KIF15), Ubiquitin Conjugating Enzyme E2 C (UBE2C), Marker of Proliferation Ki-67 (MK167), Hyaluronan-Mediated Motility Receptor (HMMR), and Cyclin B2 (CCNB2)-were identified from GEO datasets (GSE4882, GSE40275, GSE22874, GSE166720, GSE7880, GSE1987, GSE45142, GSE162102, GSE63571, and GSE23361) using the GEO2R tool. A molecular docking study involving 18 natural compounds was performed against these top 10 biomarker proteins. The compounds hesperetin (binding affinity [BA] = -10.5 kcal/mol; root mean square deviation [RMSD] = 1.569 Å) and epigallocatechin gallate (EGCG) (BA = -9.4 kcal/mol; RMSD = 0.025 Å) showed strong interactions with the TOP2A protein. To substantiate these findings, the efficacy of hesperetin and EGCG was tested in LC cells. The combination treatment had an IC₅₀ of 78 µg/mL in A549 LC cells and significantly reduced cell proliferation, migration, and TOP2A gene expression, as assessed by MTT assay, wound healing assay, and RT-PCR. The results corroborate the computational predictions and support the utility of EGCG and hesperetin in targeting LC at the cellular level.