Journal of Cancer Research and Clinical Oncology

, Volume 134, Issue 3, pp 411–419

Identifying altered gene expression in neuroblastoma cells preceding apoptosis

  • Piruz Nahreini
  • Xiang-Dong Yan
  • Cynthia P. Andreatta
  • Kedar N. Prasad
  • Neil W. Toribara
Original Paper

DOI: 10.1007/s00432-007-0303-0

Cite this article as:
Nahreini, P., Yan, XD., Andreatta, C.P. et al. J Cancer Res Clin Oncol (2008) 134: 411. doi:10.1007/s00432-007-0303-0

Abstract

Purpose

Concomitant differentiation and partial inhibition of proteasome trigger cell death in a neuroblastoma cell line (NBP2). Neither induction of differentiation nor partial inhibition of proteasome alone affects the viability of NBP2 cells. We wanted to identify genes whose expression alters under concomitant conditions and may account for cell death.

Methods

We used gel electrophoresis to analyze total genomic DNA for the detection of DNA fragmentation. Affymetrix Murine Genome U74A version 2 microarray was used to screen for ∼6,000 functionally characterized genes and ∼6,000 expressed sequence tags (ESTs). Real time PCR (RT-PCR) was performed to provide an accurate assessment of changes in gene expression.

Results

Concomitant differentiation and partial inhibition of proteasome trigger apoptosis, characterized by genomic DNA fragmentation in NBP2 cells. We found that the expression of 41 genes changed 2.5-fold or more primarily under concomitant conditions midway through apoptosis. Based on real time PCR, the expression of galectin-3, glycosylated 96, a leucine zipper protein (LRG-21), and endothelial cell activated protein C receptor (EPCR) increased between 50–500-fold, whereas the expression of Polo serine/threonine kinase, N-myc, and Histone H2A.1 decreased ranging from 8 to 37 fold. Altered expression of galectin-3, EPCR, and LRG-21 was detected as early as 2–8 h post simultaneous conditions.

Conclusion

We identified new genes that might be involved in apoptotic events in neuroblastoma cells.

Keywords

NeuroblastomaApoptosisAffymetrix microarrayProteasome inhibitionDifferentiationMitochondriaNeurodegeneration

Supplementary material

432_2007_303_MOESM1_ESM.doc (34 kb)
Real Time PCR Analysis of Fold Changes in Gene Expression (DOC 34.5 kb)

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Piruz Nahreini
    • 1
  • Xiang-Dong Yan
    • 1
  • Cynthia P. Andreatta
    • 1
  • Kedar N. Prasad
    • 2
  • Neil W. Toribara
    • 1
  1. 1.Department of Gastroenterology and Hepatology, School of MedicineUniversity of Colorado Health Sciences Center (UCHSC), Denver Health Medical Center (DHMC)DenverUSA
  2. 2.Premier Micronutrient CorporationNashvilleUSA