Therapeutic potential of BAK gene silencing in aluminum induced neural cell degeneration

J Inorg Biochem. 2009 Nov;103(11):1514-20. doi: 10.1016/j.jinorgbio.2009.06.010. Epub 2009 Aug 14.

Abstract

Previous studies have demonstrated robust BAK gene silencing via RNA interference (RNAi). To investigate whether BAK RNAi may serve as a co-therapeutic agent in neural cell death, we herein established a cell degeneration model using a human neuroblastoma cell line (SH-SY5Y) treated by aluminum (Al). Combining cell viability assays and expression analyses by QRT (quantitative real-time)-PCR and immunocytochemistry, we selected and validated the optimal small interfering RNA (siRNA) from three candidate siRNAs for the BAK gene. Our data identified siRNA1 as the most effective siRNA; the optimal concentration of the transfection agent was 10nM and the optimal incubation period was 24h. The transfection and knockdown efficiency was 93% and 58%, respectively, which closely correlated with the BAK protein expression. SH-SY5Y cells with BAK knockdown showed a clear resistance against cell death and Al-induced apoptosis. These results indicate that genetic inactivation of BAK could be an effective strategy in delaying the onset of apoptosis in Al-treated cells, and exemplify the therapeutic potential of RNAi-based methods for the treatment of neural cell degeneration.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aluminum / toxicity*
  • Apoptosis
  • Cell Line, Tumor
  • Cell Survival
  • Humans
  • Microscopy, Electron, Transmission
  • Nerve Degeneration / chemically induced*
  • Nerve Degeneration / pathology
  • Nerve Degeneration / therapy*
  • Neurons / drug effects
  • Neurons / metabolism
  • RNA Interference*
  • RNA, Small Interfering / genetics
  • Transfection
  • bcl-2 Homologous Antagonist-Killer Protein / genetics*

Substances

  • RNA, Small Interfering
  • bcl-2 Homologous Antagonist-Killer Protein
  • Aluminum