Promoting effects of sanguinarine on apoptotic gene expression in human neuroblastoma cells.

Emre Cecen,Nur Olgun,Safiye Aktas,Zekiye Altun,Pinar Ercetin

doi:10.7314/apjcp.2014.15.21.9445

Abstract

Neuroblastoma is the most common extracranial solid tumor in children. Approximately half of the affected patients are diagnosed with high-risk poor prognosis disease, and novel therapies are needed. Sanguinarine is a benzophenanthridine alkaloid which has anti-microbial, anti-oxidant and anti-inflammatory properties. The aim of this study is whether sanguinarine has in vitro apoptotic effects and which apoptotic genes might be affected in the human neuroblastoma cell lines SH-SY5Y (N-myc negative), Kelly (N-myc positive, ALK positive), and SK- N-BE(2). Cell viability was analysed with WST-1 and apoptotic cell death rates were determined using TUNEL. After RNA isolation and cDNA conversion, expression of 84 custom array genes of apoptosis was determined. Sanguinarine caused cell death in a dose dependent manner in all neuroblastoma cell lines except SK-N-BE(2) with rates of 18% in SH-SY5Y and 21% in Kelly human neuroblastoma cells. Cisplatin caused similar apoptotic cell death rates of 16% in SH-SY5Y and 23% in Kelly cells and sanguinarine-cisplatin combinations caused the same rates (18% and 20%). Sanguinarine treatment did not affect apoptototic gene expression but decreased levels of anti-apoptotic genes NOL3 and BCL2L2 in SH-SY5Y cells. Caspase and TNF related gene expression was affected by the sanguinarine-cisplatin combination in SH-SY5Y cells. The expression of regulation of apoptotic genes were increased with sanguinarine treatment in Kelly cells. From these results, we conclude that sanguinarine is a candidate agent against neuroblastoma.

Highlights

Neuroblastoma is the most common extracranial solid tumors of childhood and the most frequently diagnosed neoplasm during infancy (Missaoui et al, 2011; National cancer institute, 2014)
The aim of this study is whether sanguinarine has in vitro apoptotic effects and which apoptotic genes might be affected in the human neuroblastoma cell lines SH-SY5Y (N-myc negative), Kelly (N-myc positive, ALK positive), and SKN-BE(2)
The impact of sanguinarine on Sanguinarine-mediated apoptosis has been shown occurring through multiple pathways, including activation of nuclear factor-κB (NF-κB) (Chaturvedi et al, 1997), cell cycle arrest (Adhami et al, 2004), and mitochondrial damage (Adhami et al, 2003)

Summary

Introduction

Neuroblastoma is the most common extracranial solid tumors of childhood and the most frequently diagnosed neoplasm during infancy (Missaoui et al, 2011; National cancer institute, 2014). It accounts for more than 7% of malignancies in patients younger than 15 years and around 15% of all paediatric oncology deaths. Children younger than 18 month of age fare better than older children with the same disease stage. N-myc amplification predicts a poor outcome in all age/stage groups. Half of all patients with neuroblastoma are diagnosed with high-risk poor prognosis disease, and novel therapies are needed (Brodeur et al, 2011; Mehdiabadi et al, 2013; Wiangnon et al, 2011; Gao et al, 2014; Zhang et al, 2014)

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