Antisense bcl-2 transfection up-regulates anti-apoptotic and anti-oxidant thioredoxin in neuroblastoma cells

J Neurooncol. 2005 Mar;72(1):17-23. doi: 10.1007/s11060-004-3116-x.

Abstract

Antisense bcl-2 therapy combined with chemotherapy has been proved to be effective in various tumors. However, the role played by antisense bcl-2 therapy alone is not clear. In this study, we compared the apoptosis and the protein profiles of antisense bcl-2 transfected human neuroblastoma SK-N-MC cells to the control cells. Flow cytometric data indicated that antisense bcl-2 transfection did not lead to more extensive apoptosis in SK-N-MC cells (14.9 +/- 3.8%) than the control cells (10.3 +/- 2.3%). The above observation was confirmed by fluorescence microscopy using Hoechst 33258 staining. However, antisense bcl-2 induced changes in the expression of various proteins as shown by proteomic comparison, which included the up-regulation of the anti-apoptotic and anti-oxidant protein thioredoxin. By western blot validation, thioredoxin was found to be up-regulated by 2.9-folds with the corresponding down-regulation of Bcl-2 by 2.1-folds. The up-regulation of thioredoxin may be a compensating mechanism for cell survival in neuroblastoma when Bcl-2 expression is suppressed, and it may to some extent attenuate the effectiveness of antisense bcl-2 therapy.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / physiology*
  • Brain Neoplasms / genetics
  • Brain Neoplasms / metabolism*
  • Brain Neoplasms / therapy
  • Cell Line, Tumor
  • Cell Survival / physiology
  • DNA, Antisense / genetics
  • Gene Expression Regulation, Neoplastic
  • Genetic Therapy
  • Humans
  • Inhibitor of Apoptosis Proteins
  • Microtubule-Associated Proteins / metabolism
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism*
  • Neuroblastoma / genetics
  • Neuroblastoma / metabolism*
  • Neuroblastoma / therapy
  • Proteomics
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Proto-Oncogene Proteins c-bcl-2 / pharmacology*
  • Proto-Oncogene Proteins c-bcl-2 / physiology*
  • RNA, Messenger / analysis
  • Survivin
  • Thioredoxins / genetics
  • Thioredoxins / metabolism*
  • Transfection
  • bcl-X Protein

Substances

  • BCL2L1 protein, human
  • BIRC5 protein, human
  • DNA, Antisense
  • Inhibitor of Apoptosis Proteins
  • Microtubule-Associated Proteins
  • Neoplasm Proteins
  • Proto-Oncogene Proteins c-bcl-2
  • RNA, Messenger
  • Survivin
  • bcl-X Protein
  • Thioredoxins