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. 1988 Jun;131(3):391–397.

Detection of N-myc gene expression in neuroblastoma tumors by in situ hybridization.

P S Cohen 1, R C Seeger 1, T J Triche 1, M A Israel 1
PMCID: PMC1880701  PMID: 2454580

Abstract

The presence of N-myc DNA amplification in human neuroblastoma tumors has been shown to be an independent prognostic factor predicting rapid progression of disease. Southern blot analysis has been used previously to detect N-myc amplification in these tumors. The authors report an analysis of N-myc gene expression by in situ hybridization in 28 human neuroblastoma tumors previously studied by Southern blot analysis. In the LA-N-5 human neuroblastoma cell line known to be amplified for N-myc, reaction conditions favoring RNA-RNA hybridization yielded an optimal signal. Using these hybridization conditions, in situ hybridization analysis of N-myc expression in 28 human neuroblastoma tissues correlated perfectly with N-myc DNA amplification in these tumors as detected by Southern blot analysis. In particular, there were no tumors in which N-myc RNA expression was found by in situ hybridization analysis in the absence of DNA amplification detectable by Southern blot, nor were there tumors that had DNA amplification in the absence of RNA expression. Heterogeneity of N-myc RNA expression was observed both among cells in any given tumor area, as well as within different areas of a single tumor. N-myc expression by in situ hybridization analysis was not observed in those tumors with more neuronally differentiated, ganglioneuroma histology. It is concluded that in situ hybridization of tissue sections is as effective as Southern blot analysis of tumor cell DNA in identifying human neuroblastoma tumors in which the N-myc gene is of prognostic significance.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Brodeur G. M., Hayes F. A., Green A. A., Casper J. T., Wasson J., Wallach S., Seeger R. C. Consistent N-myc copy number in simultaneous or consecutive neuroblastoma samples from sixty individual patients. Cancer Res. 1987 Aug 15;47(16):4248–4253. [PubMed] [Google Scholar]
  2. Brodeur G. M., Seeger R. C., Schwab M., Varmus H. E., Bishop J. M. Amplification of N-myc in untreated human neuroblastomas correlates with advanced disease stage. Science. 1984 Jun 8;224(4653):1121–1124. doi: 10.1126/science.6719137. [DOI] [PubMed] [Google Scholar]
  3. Donner L., Triche T. J., Israel M. A., Seeger R. C., Reynolds C. P. A panel of monoclonal antibodies which discriminate neuroblastoma from Ewing's sarcoma, rhabdomyosarcoma, neuroepithelioma, and hematopoietic malignancies. Prog Clin Biol Res. 1985;175:347–366. [PubMed] [Google Scholar]
  4. Folks T. M., Powell D., Lightfoote M., Koenig S., Fauci A. S., Benn S., Rabson A., Daugherty D., Gendelman H. E., Hoggan M. D. Biological and biochemical characterization of a cloned Leu-3- cell surviving infection with the acquired immune deficiency syndrome retrovirus. J Exp Med. 1986 Jul 1;164(1):280–290. doi: 10.1084/jem.164.1.280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gendelman H. E., Moench T. R., Narayan O., Griffin D. E. Selection of a fixative for identifying T cell subsets, B cells, and macrophages in paraffin-embedded mouse spleen. J Immunol Methods. 1983 Dec 16;65(1-2):137–145. doi: 10.1016/0022-1759(83)90310-1. [DOI] [PubMed] [Google Scholar]
  6. Grady-Leopardi E. F., Schwab M., Ablin A. R., Rosenau W. Detection of N-myc oncogene expression in human neuroblastoma by in situ hybridization and blot analysis: relationship to clinical outcome. Cancer Res. 1986 Jun;46(6):3196–3199. [PubMed] [Google Scholar]
  7. Kohl N. E., Gee C. E., Alt F. W. Activated expression of the N-myc gene in human neuroblastomas and related tumors. Science. 1984 Dec 14;226(4680):1335–1337. doi: 10.1126/science.6505694. [DOI] [PubMed] [Google Scholar]
  8. Lee W. H., Murphree A. L., Benedict W. F. Expression and amplification of the N-myc gene in primary retinoblastoma. 1984 May 31-Jun 6Nature. 309(5967):458–460. doi: 10.1038/309458a0. [DOI] [PubMed] [Google Scholar]
  9. Maples J. A. A method for the covalent attachment of cells to glass slides for use in immunohistochemical assays. Am J Clin Pathol. 1985 Mar;83(3):356–363. doi: 10.1093/ajcp/83.3.356. [DOI] [PubMed] [Google Scholar]
  10. Nau M. M., Brooks B. J., Jr, Carney D. N., Gazdar A. F., Battey J. F., Sausville E. A., Minna J. D. Human small-cell lung cancers show amplification and expression of the N-myc gene. Proc Natl Acad Sci U S A. 1986 Feb;83(4):1092–1096. doi: 10.1073/pnas.83.4.1092. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Schwab M., Alitalo K., Klempnauer K. H., Varmus H. E., Bishop J. M., Gilbert F., Brodeur G., Goldstein M., Trent J. Amplified DNA with limited homology to myc cellular oncogene is shared by human neuroblastoma cell lines and a neuroblastoma tumour. Nature. 1983 Sep 15;305(5931):245–248. doi: 10.1038/305245a0. [DOI] [PubMed] [Google Scholar]
  12. Schwab M., Ellison J., Busch M., Rosenau W., Varmus H. E., Bishop J. M. Enhanced expression of the human gene N-myc consequent to amplification of DNA may contribute to malignant progression of neuroblastoma. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4940–4944. doi: 10.1073/pnas.81.15.4940. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Seeger R. C., Brodeur G. M., Sather H., Dalton A., Siegel S. E., Wong K. Y., Hammond D. Association of multiple copies of the N-myc oncogene with rapid progression of neuroblastomas. N Engl J Med. 1985 Oct 31;313(18):1111–1116. doi: 10.1056/NEJM198510313131802. [DOI] [PubMed] [Google Scholar]
  14. Seeger R. C., Siegel S. E., Sidell N. Neuroblastoma: clinical perspectives, monoclonal antibodies, and retinoic acid. Ann Intern Med. 1982 Dec;97(6):873–884. doi: 10.7326/0003-4819-97-6-873. [DOI] [PubMed] [Google Scholar]
  15. Thiele C. J., Reynolds C. P., Israel M. A. Decreased expression of N-myc precedes retinoic acid-induced morphological differentiation of human neuroblastoma. 1985 Jan 31-Feb 6Nature. 313(6001):404–406. doi: 10.1038/313404a0. [DOI] [PubMed] [Google Scholar]
  16. Tsuda T., Obara M., Hirano H., Gotoh S., Kubomura S., Higashi K., Kuroiwa A., Nakagawara A., Nagahara N., Shimizu K. Analysis of N-myc amplification in relation to disease stage and histologic types in human neuroblastomas. Cancer. 1987 Aug 15;60(4):820–826. doi: 10.1002/1097-0142(19870815)60:4<820::aid-cncr2820600418>3.0.co;2-y. [DOI] [PubMed] [Google Scholar]

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