Abstract

Osteosarcoma, a cancer predominantly affecting children and teenagers, has shown limited improvements in long-term survival rates despite advances in treatment, necessitating new therapeutic approaches. Natural compounds, particularly flavonoids, are being investigated for their anti-cancer properties due to their ability to modulate signaling pathways and induce apoptosis in cancer cells. This study evaluates the anti-carcinogenic effects of 6,3'-dimethoxy flavonol on MG-63 osteosarcoma cells, a p53-null model suitable for testing novel therapies, using various assays. Cell viability and proliferation were measured via MTT assay, showing dose dependent inhibition with an IC50 of 221.017 µg/ml at 48 hours. Phase-contrast microscopy revealed morphological changes consistent with apoptosis, including cell shrinkage, reduced density and cytoplasmic blebbing. Wound healing assays demonstrated significant inhibition of cell migration at 100 µg/ml and 234.12 µg/ml, highlighting its anti-metastatic potential. Acridine orange/ethidium bromide (AO/EB) staining confirmed apoptotic cell death. Real-time PCR analysis revealed an increased (Bax/Bcl-2) ratio and upregulation of p53 expression, indicating activation of the intrinsic apoptotic pathway. These findings demonstrate that 6,3'-dimethoxy flavonol effectively induces apoptosis and inhibits migration in MG-63 cells by modulating apoptotic markers and signaling pathways. The results suggest its potential as a therapeutic agent for osteosarcoma. Future studies should explore itsinvivo efficacy, possible synergistic effects in combination therapies, and its mechanisms in p53-positive cell lines. This evidence underscores the promise of flavonoid based interventions in cancer treatment.