Antiproliferative and apoptotic effect of LY2090314, a GSK-3 inhibitor, in neuroblastoma in vitro

• Published in: BMC cancer Vol. 18; no. 1; pp. 560 - 8
• Main Authors: Kunnimalaiyaan, Selvi, Schwartz, Victoriana K., Jackson, Iris Alao, Clark Gamblin, T., Kunnimalaiyaan, Muthusamy
• Format: Journal Article
• Language: English
• Published: London BioMed Central 11.05.2018; BioMed Central Ltd; BMC
• Subjects: Analysis; Antineoplastic Agents - pharmacology; Antineoplastic Agents - therapeutic use; Apoptosis; Apoptosis - drug effects; Biomedical and Life Sciences; Biomedicine; Cancer Research; Care and treatment; Cell Line, Tumor; Cell proliferation; Cell Proliferation - drug effects; Drug Screening Assays, Antitumor; Experimental therapeutics and drug development; Glycogen Synthase Kinase 3 - antagonists & inhibitors; Glycogen Synthase Kinase 3 - metabolism; GSK-3 alpha; GSK-3 beta; GSK-3 inhibitor; Health Promotion and Disease Prevention; Heterocyclic Compounds, 3-Ring - pharmacology; Heterocyclic Compounds, 3-Ring - therapeutic use; Humans; Maleimides - pharmacology; Maleimides - therapeutic use; Medicine/Public Health; N-Myc Proto-Oncogene Protein - genetics; Neuroblastoma; Neuroblastoma - drug therapy; Neuroblastoma - genetics; Oncology; Phosphorylation - drug effects; Research Article; Risk factors; Surgical Oncology; GSK-3 alpha; Neuroblastoma; GSK-3 inhibitor; GSK-3 beta; Apoptosis
• ISSN: 1471-2407; 1471-2407
• DOI: 10.1186/s12885-018-4474-7

Background Neuroblastoma (NB) is a devastating disease. Despite recent advances in the treatment of NB, about 60% of high-risk NB will have relapse and therefore long-term event free survival is very minimal. We have reported that targeting glycogen synthase kinase-3 (GSK-3) may be a potential strategy to treat NB. Consequently, investigating LY2090314, a clinically relevant GSK-3 inhibitor, on NB cellular proliferation and may be beneficial for NB treatment. Methods The effect of LY2090314 was compared with a previously studied GSK-3 inhibitor, Tideglusib. Colorimetric, clonogenic, and live-cell image confluency assays were used to study the proliferative effect of LY2090314 on NB cell lines (NGP, SK-N-AS, and SH-SY-5Y). 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 a 20 nM concentration in NGP, SK-N-AS, and SH-SY-5Y cells. Western blot 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 a reduction in the anti-apoptotic protein, survivin. Further, treatment significantly reduced the level of cyclin D1, a key regulatory protein of the 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 in vitro. To our knowledge, this is the first study to look at the effect of LY2090314 in NB cell lines. These results indicate that GSK-3 may be a therapeutic target for NB and provide rationale for further preclinical analysis using LY2090314.