Evaluation of Cytotoxic Activity and Apoptosis-Inducing Potential of 5,6,7-Trihydroxyflavone against Breast Cancer and Cervical Cancer Cell Lines

Abstract

Background: Breast cancer and cervical cancer are the leading causes of mortality in women worldwide, necessitating ongoing efforts to develop effective treatments. Anticancer drug development is an important aspect of cancer research, with the goal of discovering and designing compounds that target specific aspects of cancer biology while causing minimal harm to normal cells. Compounds that interact with DNA are an intriguing class of chemotherapeutic drugs used to treat cancer. This study investigated the anticancer effect of 5,6,7-Trihydroxyflavone (THF) on breast cancer and cervical cancer cells. Methods: The anticancer potential of 5,6,7-Trihydroxyflavone (THF) against two human breast cancer cell lines (the MDA-MB231 cell line (HTB-26™) and the MCF-7 cell line (HTB-22™)) and one human cervical cancer cell line (the HeLa cell line and the CRM-CCL-2™) was explored using in-vitro cell based assays including 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay, Lactate dehydrogenase release and Caspase-3 activity followed by the Herring sperm DNA (HS-DNA) binding UV-visible spectrophotometric assay. For in-silico investigations, Molecular docking studies (using P53, Caspase, and nuclear factor-kappa B (NF-κB)) and molecular dynamic simulation studies were performed followed by the density functional theorys (DFTs) calculations. Results: Significant growth inhibition (GI₅₀) values of 11.1 ± 1.06, 3.23 ± 0.81, and 6.14 ± 0.96 μM were found against HeLa, MDA-MB231, and MCF-7 cells, respectively. From the LDH-release assay and Caspase-3 assay, THF was found more potential towards MDA-MB231 cells. The results showed that, compared with control, the percentage LDH release and Caspase-3 activities of THF were found statistically significant (increased) when compared between the two different concentrations (p < 0.05) and were found highly significant as p < 0.001, compared to control. THF was also discovered to bind to DNA via groove binding, with both electrostatic and non-electrostatic interactions contributing to the free energy of the binding. At the end, THFs remarkable cytotoxic potential was demonstrated by molecular docking and molecular dynamics simulations. Conclusion: These findings suggest the potential application of THF that this could be the hypothetical compound, which would need extensive preclinical and clinical testing before being considered a viable treatment option.