Detection of N-myc gene amplification by fluorescence in situ hybridization. Diagnostic utility for neuroblastoma.
We assessed fluorescence in situ hybridization (FISH) as an alternative to Southern blot analysis for determination of N-myc gene amplification in neuroblastoma. In the 44 pediatric solid tumor cell lines examined (20 neuroblastomas), the mean number of N-myc copies determined by FISH correlated closely with Southern blot results. There was wide intercellular variability in gene copy number in tumors that had evidence of amplification; however, tumors judged to be non-amplified completely lacked any cells with high N-myc copy number. FISH provided reliable estimates of N-myc amplification in 12 clinical samples even when the percentage of tumor was low. The other advantages of FISH over Southern blot analysis were speed and technical simplicity, ability to discern heterogeneous gene amplification among tumor cells in the same specimen, and capacity to determine the source of the amplified N-myc signal, whether extrachromosomal double-minute chromosomes, expanded intrachromosomal regions, or chromosome 2 aneuploidy. We conclude that FISH would refine the analysis of N-myc amplification in neuroblastoma and thus improve the assignment of patients to prognostic groups based on this unfavorable risk factor.
Shapiro
D N
DN
Department of Experimental Oncology, St. Jude Children's Research Hospital, Memphis, TN 38105.
Valentine
M B
MB
Rowe
S T
ST
Sinclair
A E
AE
Sublett
J E
JE
Roberts
W M
WM
Look
A T
AT
D015139:Blotting, Southern; D002675:Child, Preschool; D002875:Chromosomes; D003720:Densitometry; D005784:Gene Amplification; D016259:Genes, myc; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007223:Infant; D009447:Neuroblastoma / Q000175:diagnosis* / Q000235:genetics; D013194:Staining and Labeling; D014407:Tumor Cells, Cultured