PMID- 14694373
OWN - NLM
STAT- MEDLINE
DCOM- 20040226
LR  - 20190710
IS  - 1531-5037 (Electronic)
IS  - 0022-3468 (Linking)
VI  - 39
IP  - 1
DP  - 2004 Jan
TI  - Clinical significance of a highly sensitive analysis for gene dosage and the 
      expression level of MYCN in neuroblastoma.
PG  - 63-8
AB  - BACKGROUND: The amplification of the MYCN gene is one of the most powerful 
      adverse prognosis factors in neuroblastoma, but the clinical significance of an 
      enhanced expression of MYCN remains controversial. To reassess the clinical 
      implications of MYCN amplification and expression in neuroblastoma, the status of 
      amplification and the expression level of the MYCN gene of primary neuroblastoma 
      samples were analyzed using highly sensitive analyses. METHODS: Using a 
      quantitative polymerase chain reaction (PCR) method (TaqMan), the gene dosages 
      (MYCN/p53) of 66 primary neuroblastoma samples were determined. In all 66 
      samples, the status of MYCN amplification has been determined previously by the 
      Southern blotting method. Of the 54 samples with a single copy of MYCN based on 
      the Southern blotting method, 23 samples were analyzed for MYCN amplification 
      using the fluorescence in situ hybridization (FISH) method. The expression levels 
      (MYCN/GAPDH) of 56 samples were determined by a quantitative reverse 
      transcriptase (RT)-PCR method. RESULTS: Of the 54 samples with a single copy of 
      MYCN based on the Southern blotting method, 46 samples showed MYCN gene dosages 
      of less than 2.0, whereas the remaining 8 samples with dosages of more than 2.0 
      were tumors from patients with advanced-stage disease. The results of FISH 
      supported the fact that these 8 samples contained a small number of 
      MYCN-amplified cells. The cases of MYCN gene dosages of more than 2.0 were 
      significantly associated with all other unfavorable prognostic factors (an age of 
      >1 year at diagnosis [P <.0001], nonmass screening [P =.0003], advanced stage [P 
      <.0001], diploid or tetraploid [P <.0001], and a Shimada unfavorable histology [P 
      <.0001]). MYCN gene dosages of more than 2.0 were significantly associated with a 
      high expression of MYCN (P =.0459). However, the expression level of MYCN was not 
      significantly associated with any other prognostic factors. CONCLUSIONS: 
      Quantitative PCR may thus be a useful modality for performing a highly sensitive 
      and accurate assessment of the amplification and expression levels of the MYCN 
      gene. In particular, the combination of the quantitative PCR system and the FISH 
      method is considered to be a highly effective method for evaluating the status of 
      MYCN amplification. In this highly sensitive analysis, MYCN amplification 
      (MYCN/p53 > or = 2.0) was reconfirmed to be a strongly unfavorable factor, 
      whereas the expression level of MYCN does not appear to be an independently 
      significant prognosis factor.
FAU - Tanaka, Shinji
AU  - Tanaka S
AD  - Department of Pediatric Surgery, Graduate School of Medical Sciences, Kyushu 
      University, Fukuoka, Japan.
FAU - Tajiri, Tatsuro
AU  - Tajiri T
FAU - Noguchi, Shin-ichi
AU  - Noguchi S
FAU - Shono, Kumiko
AU  - Shono K
FAU - Ihara, Kenji
AU  - Ihara K
FAU - Hara, Toshiro
AU  - Hara T
FAU - Suita, Sachiyo
AU  - Suita S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Pediatr Surg
JT  - Journal of pediatric surgery
JID - 0052631
RN  - 0 (MYCN protein, human)
RN  - 0 (N-Myc Proto-Oncogene Protein)
RN  - 0 (Nuclear Proteins)
RN  - 0 (Oncogene Proteins)
RN  - 63231-63-0 (RNA)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Child
MH  - Child, Preschool
MH  - DNA/isolation & purification
MH  - Female
MH  - Gene Amplification
MH  - *Gene Dosage
MH  - *Gene Expression
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Infant
MH  - Male
MH  - N-Myc Proto-Oncogene Protein
MH  - Neuroblastoma/*genetics/metabolism
MH  - Nuclear Proteins/*genetics/metabolism
MH  - Oncogene Proteins/*genetics/metabolism
MH  - Polymerase Chain Reaction
MH  - Prognosis
MH  - RNA/genetics/isolation & purification/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction
EDAT- 2003/12/25 05:00
MHDA- 2004/02/27 05:00
CRDT- 2003/12/25 05:00
PHST- 2003/12/25 05:00 [pubmed]
PHST- 2004/02/27 05:00 [medline]
PHST- 2003/12/25 05:00 [entrez]
AID - S0022346803007073 [pii]
AID - 10.1016/j.jpedsurg.2003.09.015 [doi]
PST - ppublish
SO  - J Pediatr Surg. 2004 Jan;39(1):63-8. doi: 10.1016/j.jpedsurg.2003.09.015.