Promoting Effects of Sanguinarine on Apoptotic Gene Expression in Human Neuroblastoma Cells

Abstract

Neuroblastoma is the most common extracranial solid tumor in children. Approximately half of the affectedpatients are diagnosed with high-risk poor prognosis disease, and novel therapies are needed. Sanguinarine is abenzophenanthridine alkaloid which has anti-microbial, anti-oxidant and anti-inflammatory properties. The aimof this study is whether sanguinarine has in vitro apoptotic effects and which apoptotic genes might be affected inthe human neuroblastoma cell lines SH-SY5Y (N-myc negative), Kelly (N-myc positive, ALK positive), and SKN-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 apoptoticcell death rates of 16% in SH-SY5Y and 23% in Kelly cells and sanguinarine-cisplatin combinations caused thesame rates (18% and 20%). Sanguinarine treatment did not affect apoptototic gene expression but decreasedlevels of anti-apoptotic genes NOL3 and BCL2L2 in SH-SY5Y cells. Caspase and TNF related gene expressionwas affected by the sanguinarine-cisplatin combination in SH-SY5Y cells. The expression of regulation ofapoptotic genes were increased with sanguinarine treatment in Kelly cells. From these results, we conclude thatsanguinarine is a candidate agent against neuroblastoma.