Stable Expression of Short Interfering RNA for DT-Diaphorase Induces Neurotoxicity

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• 作者：Jorge Lozano ; Patricia Muoz ; Beston F. Nore ; Susan LeDoux ; Juan Segura-Aguilar
• 刊名：Chemical Research in Toxicology
• 出版年：2010
• 出版时间：September 20, 2010
• 年：2010
• 卷：23
• 期：9
• 页码：1492-1496
• 全文大小：176K
• 年卷期：v.23,no.9(September 20, 2010)
• ISSN：1520-5010

文摘

DT-Diaphorase has been proposed to play a neuroprotective role in dopaminergic neurons by preventing aminochrome neurotoxicity. There are several studies supporting this idea, but in all studies, we used dicoumarol, an inhibitor of DT-diaphorase. We have designed and developed two siRNA to silence the expression of DT-diaphorase to study its role in aminochrome metabolism. We transduced RCSN-3 cells with retroviral particles containing a pRetroSuper plasmid coding a siRNA for DT-diaphorase. The cells selected in the presence of puromycin generated a stable cell line RCSN-3Nq6 and RCSN-3Nq7 with low expression of DT-diaphorase (27% and 33% of wild type, respectively). A significant cell death was observed in RCSN-3 cells expressing siRNA Nq6 and Nq7 for DT-diaphorase when were incubated with 100 μM aminochrome during 48 (4- and 3.5-fold, respectively; P < 0.01). These results support the protective role of DT-diaphorase against aminochrome neurotoxicity in dopaminergic neurons containing neuromelanin and show that Nq6 and Nq7 siRNA are very useful tools to study the role of DT-diaphorase in aminochrome metabolism.