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Inhibitory Effects of Hesperetin and EGCG in the Lung Cancer Progression involves impairment in TOP2A gene expression
Rangaraj Kaviyaprabha1, Thandaserry Vasudevan Miji1,2, Unni Apsara1
1Centre for Bioinformatics, Department of Biochemistry, Karpagam Academy of Higher Education, Tamil Nadu, Coimbatore, 641021, India.
Abstract:
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.
Insights
This study identifies key lung cancer biomarkers and evaluates natural compounds, finding hesperetin and EGCG effectively target DNA Topoisomerase II Alpha (TOP2A) in lung cancer cells, reducing proliferation and migration.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Lung cancer (LC) is a leading cause of cancer mortality, with Non-Small Cell Lung Cancer (NSCLC) treatments facing challenges due to drug resistance.
- Identifying novel therapeutic targets and effective compounds is crucial for improving treatment outcomes and overcoming resistance mechanisms in LC.
Purpose of the Study:
- To identify key biomarkers associated with lung cancer progression and drug resistance.
- To evaluate the potential of natural compounds as therapeutic agents targeting identified lung cancer biomarkers, specifically DNA Topoisomerase II Alpha (TOP2A).
Main Methods:
- Utilized GEO datasets and GEO2R tool to identify ten key lung cancer biomarkers.
- Performed in silico molecular docking of 18 natural compounds against identified biomarker proteins.
- Conducted in vitro validation of promising compounds (hesperetin and epigallocatechin gallate - EGCG) in A549 lung cancer cells using MTT assay, wound healing assay, and RT-PCR.
Main Results:
- Identified ten key LC biomarkers, including ASPM, BUB1, CENPF, TOP2A, TPX2, KIF15, UBE2C, MK167, HMMR, and CCNB2.
- Hesperetin and EGCG demonstrated strong binding affinity to the TOP2A protein in molecular docking studies.
- Combined hesperetin and EGCG treatment significantly inhibited proliferation and migration, and reduced TOP2A gene expression in A549 cells.
Conclusions:
- Hesperetin and EGCG show significant potential as therapeutic agents for lung cancer by targeting TOP2A.
- The study validates the in silico findings through in vitro experiments, supporting the use of these natural compounds in targeting lung cancer at the cellular level.
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