Use of RNA interference to elucidate the effect of MYCN on cell cycle in neuroblastoma

Pediatr Blood Cancer. 2008 Feb;50(2):208-12. doi: 10.1002/pbc.21195.


Background: MYCN amplification marks poor prognosis in neuroblastoma (NB) tumors. In evaluating the mechanisms by which retinoic acid (RA) or nerve growth factor (NGF) decrease cell number in MYCN amplified NB cells, we have identified a number of proteins whose expression either decreases (E2F, CDC2, CDK6, cyclin dependent kinase activity) or increases (p27) in association with a decrease in MYCN expression. However, it was still unclear which were MYCN dependent effects or not.

Procedure: This study aimed to determine which changes in cell cycle gene expression are modulated as a consequence of the decrease in MYCN. We silenced MYCN expression using siRNA targeted to the coding region of MYCN. Then, by using siRNA transient transfections, we analyzed the change of cell cycle related genes and cell cycle in MYCN amplified NB cell lines.

Results: We demonstrate that expression of MYCN can be suppressed by almost 60% in MYCN amplified NB cell using siRNAs targeted to MYCN. Functionally, the decrease in MYCN leads to a decrease in cells in the S-phase of the cell cycle. Decreases in MYCN are associated with decreases in E2F1-2 and ID2 along with increases in p27 protein levels by post-transcriptional modification. Moreover, we find that a decrease in MYCN is accompanied by a decrease in cdk6 mRNA and protein expression.

Conclusions: These results show that E2F and ID2 expression is associated with MYCN regulation and that cdk6 is a possible new transcriptional target of MYCN.

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology
  • Cell Cycle / genetics
  • Cell Cycle Proteins / biosynthesis
  • Cell Cycle Proteins / genetics
  • Cell Growth Processes / genetics
  • Cell Line, Tumor
  • Gene Silencing
  • Humans
  • Mice
  • N-Myc Proto-Oncogene Protein
  • NIH 3T3 Cells
  • Neuroblastoma / genetics*
  • Neuroblastoma / metabolism
  • Neuroblastoma / pathology*
  • Nuclear Proteins / biosynthesis
  • Nuclear Proteins / deficiency
  • Nuclear Proteins / genetics*
  • Oncogene Proteins / biosynthesis
  • Oncogene Proteins / deficiency
  • Oncogene Proteins / genetics*
  • RNA Interference*
  • RNA, Small Interfering / genetics
  • Transfection
  • Tretinoin / pharmacology


  • Antineoplastic Agents
  • Cell Cycle Proteins
  • MYCN protein, human
  • N-Myc Proto-Oncogene Protein
  • Nuclear Proteins
  • Oncogene Proteins
  • RNA, Small Interfering
  • Tretinoin