Image-Based and Molecular Profiling of Thymol and Thymoquinone as Dual VEGFR-2/COX-2 Inhibitors in Lung and Breast Cancer

Lung and breast cancers are major causes of cancerrelated deaths, necessitating the development of novel therapies. Breast cancer metastasis to the lungs is driven by vascular endothelial growth factor receptor-2 (VEGFR-2)-mediated angiogenesis and cyclooxygenase-2 (COX-2) induced inflammation, which create a favorable microenvironment for tumor growth. This study investigates the dual-target potential of thymol (THY) and thymoquinone (TQ) to inhibit VEGFR-2 and COX-2. By using carboplatin (CP) as a positive control, we aimed to evaluate this combinatorial approach. The findings could pave the way for innovative treatments for metastatic breast cancer. In silico studies using ADMET analysis and molecular docking were performed to assess the drug-likeness and binding affinities of THY and TQ toward VEGFR-2 and COX-2 where both compounds demonstrated superior binding profiles compared to CP. In vitro studies showed that the combination of THY+TQ exerted significant dose-dependent cytotoxicity in lung and breast cancer cell lines. Phase contrast microscopy confirmed apoptotic morphology in all THY+TQ treated cell lines, while cell cycle analysis revealed a marked arrest at the G0/G1 phase. Wound healing assays demonstrated a substantial reduction in cell migration in combination THY+TQ treatment. Furthermore, realtime PCR analysis showed downregulation of VEGFR-2 and COX-2 expression. These results highlight the potential of this combination therapy as a promising adjunct to conventional chemotherapy. Further in vivo studies are required to validate these findings.