Abstract
OBJECTIVES: The present study aimed to assess the antiproliferative and pro-apoptotic effects of hinokitiol in osteosarcoma cells via in vitro and in silico targeting of glycogen synthase kinase 3β (GSK3β). MATERIALS AND METHODS: The (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used to evaluate the cytotoxic potential of hinokitiol in osteosarcoma cells. Various concentrations of hinokitiol (5, 10, 20, 40, 60, and 80 μg/mL) were tested, and the half-maximal inhibitory concentration (IC(50)) was calculated. Cell morphology, migration (scratch assay), and gene expression analysis using real-time polymerase chain reaction for pro-apoptotic studies were conducted, with the IC(50) dose of hinokitiol utilized in all these experiments. Additionally the anti-proliferative effect of hinokitiol on GSK3β was also examined using in silico and gene expression methods. RESULTS: Hinokitiol significantly (p < 0.05) and dose-dependently decreased the viability of MG-63 cells, with an IC(50) value of 40 μg/mL. Cell morphology study revealed cellular shrinkage and reduced cell density. The scratch assay revealed anti-migratory activity, while gene expression studies indicated pro-apoptotic effects, including significant (p < 0.05) upregulation of BAX and down-regulation of BCL-2 and GSK3β. Bonding interactions were also observed with GSK3β and atomic contact energy of -5.69 kcal/mol. CONCLUSION: According to the current study findings, hinokitiol prevented Morphological study of the effects of hinokitiol on osteosarcoma cells from proliferating, migrating, and induced apoptosis by upregulating BAX (a pro-apoptotic signal) expression and downregulating BCL-2 (anti-apoptotic signal) expression in osteosarcoma cells. In silico findings of hinokitiol showed a significant bonding interaction with GSK3β and its downregulated gene expression probably prevented cancer cell survival.