Original Article

In silico and In vitro Evaluation of Cytotoxic Potential of Hinokitiol against Osteosarcoma by targeting Glycogen Synthase Kinase 3β


  • Binoy Varghese Cheriyan
  • Punithavel Srinivasan
  • George J Push
  • Karthik K K
  • Prasanna Bharathi Sainath
  • Anandakumar Shanmugam
  • Anitha Roy
  • Karthikeyan Elumalai

Received Date: 10.06.2023 Accepted Date: 23.12.2023 Turk J Pharm Sci 0;0(0):0-0 [e-Pub]


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.

Material and Methods:

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.

Keywords: Hinokitiol, Osteosarcoma, MTT assay, in silico, Glycogen synthase kinase 3β