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

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

Chan-Wook Woo  1 , Fei Tan, Hope Cassano, Junghwa Lee, Kwang Chul Lee, Carol J Thiele
Affiliations

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

Chan-Wook Woo et al. Pediatr Blood Cancer. 2008 Feb.

Abstract

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.

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