Objective: Our objective is to test the hypothesis that LY2090314, a clinically relevant GSK-3 inhibitor, will effectively inhibit neuroblastoma cellular proliferation. Methods: Effect of LY2090314 was compared with... [ view full abstract ]
Objective:
Our objective is to test the hypothesis that LY2090314, a clinically relevant GSK-3 inhibitor, will effectively inhibit neuroblastoma cellular proliferation.
Methods: Effect of LY2090314 was compared with previously studied GSK-3 inhibitor, Tideglusib. Colorimetric, colonogenic, and live-cell image confluency assays were used to study the proliferative effect of LY2090314 on NB cell lines (NGP, SK-N-AS, and Neuro2A). Western blotting and caspase glo assay were performed to determine the mechanistic function of LY2090314 in NB cell lines.
Results: LY2090314 treatment exhibited significant growth reduction starting at 20nM concentration in NGP compared to SK-N-AS and Neuro2A cells (5µM). Western analysis indicated that growth suppression was due to apoptosis as evidenced by an increase in pro-apoptotic markers cleaved PARP and cleaved caspase-3 and reduction in anti-apoptotic proteins, survivin. Furthermore, treatment significantly reduced the level of cyclinD1, a key regulatory protein for cell cycle and apoptosis. Functionally, this was confirmed by an increase in caspase activity. LY2090314 treatment reduced the expression levels of phosphorylated GSK-3 proteins and increased the stability of β-catenin in these cells.
Conclusions: LY2090314 effectively reduces growth of both human MYCN amplified and non-amplified NB cell lines as well as mouse derived NB cells in vitro. To our knowledge, this is the first study on the effect of LY2090314 in NB cell lines in vitro and provides rationale for further preclinical analysis
