ABSTRACT

The current study aimed to assess the anti-proliferative and pro-apoptotic effects of hinokitiol on osteosarcoma cells by in vitro and in silico targeting of glycogen synthase kinase 3β.

The MTT assay was used to evaluate the cytotoxic potential of hinokitiol in osteosarcoma cells, Hinokitiol was utilized in a variety of concentrations (5, 10, 20, 40, 60, 80μg/ml), and inhibitory concentration IC₅₀ dose were calculated. Cell morphology, migration (scratch assay), and gene regulation for expression (RT-PCR) for pro-apoptosis study were also conducted and IC₅₀ dose was considered for the aforesaid studies. The role of hinokitiol's anti-proliferative effects on glycogen synthase kinase-3β was also examined using in silico and its gene expression methods.

Hinokitiol dose-dependently decreased the viability of MG-63 cells with an IC50 of 40μg/ml. Cell morphology study finding revealed cellular shrinkage and reduced cell density, Scratch assay revealed it had anti-migratory activity, and pro-apoptotic property of target genes was revealed in the gene expression study. Bonding interactions were also observed with glycogen synthase kinase-3β and atomic contact energy observed was -5.69kcal/mol

According to the current study's findings, Hinokitiol prevented MG63 cells from proliferating, migrating and induced apoptosis effect via upregulation of BAX (a proapoptotic signal) expression downregulation of BCL-2 (antiapoptotic signal) expression, in osteosarcoma cells. In silico findings of hinokitiol showed significant bonding interaction with glycogen synthase kinase 3 β and its downregulated gene expression probably preventing cancer cell survival.