Repositioning of Metformin: Anticancer Agent for Hypoxic Neuroblastoma Cells

Abstract

Objectives:Neuroblastoma is an extracranial solid tumor of early childhood that has a hypoxic environment. VEGF and HIFs molecules play a role in adaptation to this microenvironment. Hypoxic microenvironment leads to poor prognosis and inadequate treatment of neuroblastoma. Metformin has been shown to inhibit tumor growth, might be a potential chemotherapeutic agent. The anti-cancer activity of Metformin on SH-SY5Y cells are not fully elucidated. The aim of this study is to determine the anti-cancer effect of Metformin on SH-SY5Y cells and to elucidate its molecular action mechanism in hypoxia/normoxia. Patients and Methods:SH-SY5Y cells were exposed to increasing doses of Metformin. The viability of SH-SY5Y cells was evaluated using the real time xCELLigence RTCA system. Migration of SH-SY5Y cells was determined using wound healing. The effect of metformin on mRNA and protein expression levels was evaluated using Real-time PCR and Western Blot, respectively. Results:Metformin was observed to significantly reduce the viability of SH-SY5Y cells. Metformin treatment reduced migration of SH-SY5Y cells. In addition, Metformin treatment significantly reduced mRNA expression of HIF-1α, PDK-1 and VEGF-A in SH-SY5Y cells under normoxia and hypoxia. In hypoxia condition, protein expression of HIF-1α and VEGF-A decreased after Metformin administration on SH-SY5Y cells. Protein expression of PDK-1 was observed to decrease in both normoxia and hypoxia conditions on SH-SY5Y cells. Conclusion:In this study, the anti-cancer effect of Metformin on SH-SY5Y cells was determined. Metformin has been observed as inhibitor of migration in SH-SY5Y. Metformin treatment has been shown to reduce gene and protein levels of HIF-1α and target molecules in neuroblastoma cells.

Keywords

• neuroblastoma, metformin, hypoxia, HIF-1α, VEGF-A, PDK-1

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