Skip to main content
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1990 Feb;10(2):823–829. doi: 10.1128/mcb.10.2.823

Excision of N-myc from chromosome 2 in human neuroblastoma cells containing amplified N-myc sequences.

J D Hunt 1, M Valentine 1, A Tereba 1
PMCID: PMC360884  PMID: 2405257

Abstract

Amplification of one of three growth-stimulating myc genes is a common method by which many tumor types gain a proliferative advantage. In metastatic human neuroblastoma, the amplification of the N-myc locus, located on chromosome 2, is a dominant feature of this usually fatal pediatric cancer. Of the many models proposed to explain this amplification, all incorporate as the initial step either disproportionate overreplication of the chromosomal site or recombination across a loop structure. The original locus is retained within the chromosome in the overreplication models but is excised in the recombination models. To test these models, we have used somatic cell hybrids to separate and analyze the chromosomes 2 from a neuroblastoma cell line containing in vivo amplified N-myc. Our results demonstrate that N-myc is excised from one of the chromosomes, suggesting that deletion is a requisite part of gene amplification in a naturally occurring system.

Full text

PDF
823

Images in this article

Selected References

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

  1. Brodeur G. M., Green A. A., Hayes F. A., Williams K. J., Williams D. L., Tsiatis A. A. Cytogenetic features of human neuroblastomas and cell lines. Cancer Res. 1981 Nov;41(11 Pt 1):4678–4686. [PubMed] [Google Scholar]
  2. Carroll S. M., DeRose M. L., Gaudray P., Moore C. M., Needham-Vandevanter D. R., Von Hoff D. D., Wahl G. M. Double minute chromosomes can be produced from precursors derived from a chromosomal deletion. Mol Cell Biol. 1988 Apr;8(4):1525–1533. doi: 10.1128/mcb.8.4.1525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cohn S. L., Salwen H., Herst C. V., Maurer H. S., Nieder M. L., Morgan E. R., Rosen S. T. Single copies of the N-myc oncogene in neuroblastomas from children presenting with the syndrome of opsoclonus-myoclonus. Cancer. 1988 Aug 15;62(4):723–726. doi: 10.1002/1097-0142(19880815)62:4<723::aid-cncr2820620413>3.0.co;2-u. [DOI] [PubMed] [Google Scholar]
  4. Collins S., Groudine M. Amplification of endogenous myc-related DNA sequences in a human myeloid leukaemia cell line. Nature. 1982 Aug 12;298(5875):679–681. doi: 10.1038/298679a0. [DOI] [PubMed] [Google Scholar]
  5. Cowell J. K. Double minutes and homogeneously staining regions: gene amplification in mammalian cells. Annu Rev Genet. 1982;16:21–59. doi: 10.1146/annurev.ge.16.120182.000321. [DOI] [PubMed] [Google Scholar]
  6. Derom C., Bakker E., Vlietinck R., Derom R., Van den Berghe H., Thiery M., Pearson P. Zygosity determination in newborn twins using DNA variants. J Med Genet. 1985 Aug;22(4):279–282. doi: 10.1136/jmg.22.4.279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  8. Futcher A. B. Copy number amplification of the 2 micron circle plasmid of Saccharomyces cerevisiae. J Theor Biol. 1986 Mar 21;119(2):197–204. doi: 10.1016/s0022-5193(86)80074-1. [DOI] [PubMed] [Google Scholar]
  9. Gillin F. D., Roufa D. J., Beaudet A. L., Caskey C. T. 8-Azaguanine resistance in mammalian cells. I. Hypoxanthine-guanine phosphoribosyltransferase. Genetics. 1972 Oct;72(2):239–252. doi: 10.1093/genetics/72.2.239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hamlin J. L., Milbrandt J. D., Heintz N. H., Azizkhan J. C. DNA sequence amplification in mammalian cells. Int Rev Cytol. 1984;90:31–82. doi: 10.1016/s0074-7696(08)61487-4. [DOI] [PubMed] [Google Scholar]
  11. Hunt J. D., Goren M. P., Tereba A. Agarose gel electrophoresis in isozyme separation and visualization. Biochem Genet. 1989 Dec;27(11-12):647–654. doi: 10.1007/BF02396057. [DOI] [PubMed] [Google Scholar]
  12. Jakobovits A., Schwab M., Bishop J. M., Martin G. R. Expression of N-myc in teratocarcinoma stem cells and mouse embryos. Nature. 1985 Nov 14;318(6042):188–191. doi: 10.1038/318188a0. [DOI] [PubMed] [Google Scholar]
  13. Kaufman R. J., Sharp P. A., Latt S. A. Evolution of chromosomal regions containing transfected and amplified dihydrofolate reductase sequences. Mol Cell Biol. 1983 Apr;3(4):699–711. doi: 10.1128/mcb.3.4.699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Kohl N. E., Legouy E., DePinho R. A., Nisen P. D., Smith R. K., Gee C. E., Alt F. W. Human N-myc is closely related in organization and nucleotide sequence to c-myc. Nature. 1986 Jan 2;319(6048):73–77. doi: 10.1038/319073a0. [DOI] [PubMed] [Google Scholar]
  16. McCLINTOCK B. Chromosome organization and genic expression. Cold Spring Harb Symp Quant Biol. 1951;16:13–47. doi: 10.1101/sqb.1951.016.01.004. [DOI] [PubMed] [Google Scholar]
  17. Nau M. M., Brooks B. J., Battey J., Sausville E., Gazdar A. F., Kirsch I. R., McBride O. W., Bertness V., Hollis G. F., Minna J. D. L-myc, a new myc-related gene amplified and expressed in human small cell lung cancer. Nature. 1985 Nov 7;318(6041):69–73. doi: 10.1038/318069a0. [DOI] [PubMed] [Google Scholar]
  18. Passananti C., Davies B., Ford M., Fried M. Structure of an inverted duplication formed as a first step in a gene amplification event: implications for a model of gene amplification. EMBO J. 1987 Jun;6(6):1697–1703. doi: 10.1002/j.1460-2075.1987.tb02420.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ritke M. K., Shah R., Valentine M., Douglass E. C., Tereba A. Molecular analysis of chromosome 1 abnormalities in neuroblastoma. Cytogenet Cell Genet. 1989;50(2-3):84–90. doi: 10.1159/000132729. [DOI] [PubMed] [Google Scholar]
  20. Ruiz J. C., Wahl G. M. Formation of an inverted duplication can be an initial step in gene amplification. Mol Cell Biol. 1988 Oct;8(10):4302–4313. doi: 10.1128/mcb.8.10.4302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Saiki R. K., Scharf S., Faloona F., Mullis K. B., Horn G. T., Erlich H. A., Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. doi: 10.1126/science.2999980. [DOI] [PubMed] [Google Scholar]
  22. Schimke R. T. Gene amplification, drug resistance, and cancer. Cancer Res. 1984 May;44(5):1735–1742. [PubMed] [Google Scholar]
  23. 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]
  24. Schwab M., Ramsay G., Alitalo K., Varmus H. E., Bishop J. M., Martinsson T., Levan G., Levan A. Amplification and enhanced expression of the c-myc oncogene in mouse SEWA tumour cells. Nature. 1985 May 23;315(6017):345–347. doi: 10.1038/315345a0. [DOI] [PubMed] [Google Scholar]
  25. Schwab M., Varmus H. E., Bishop J. M., Grzeschik K. H., Naylor S. L., Sakaguchi A. Y., Brodeur G., Trent J. Chromosome localization in normal human cells and neuroblastomas of a gene related to c-myc. Nature. 1984 Mar 15;308(5956):288–291. doi: 10.1038/308288a0. [DOI] [PubMed] [Google Scholar]
  26. Schwab M., Varmus H. E., Bishop J. M. Human N-myc gene contributes to neoplastic transformation of mammalian cells in culture. Nature. 1985 Jul 11;316(6024):160–162. doi: 10.1038/316160a0. [DOI] [PubMed] [Google Scholar]
  27. Skolnick M. H., Francke U. Report of the committee on human gene mapping by recombinant DNA techniques. Oslo Conference (1981): Sixth International Workshop on Human Gene Mapping. Cytogenet Cell Genet. 1982;32(1-4):194–204. doi: 10.1159/000131699. [DOI] [PubMed] [Google Scholar]
  28. Small M. B., Hay N., Schwab M., Bishop J. M. Neoplastic transformation by the human gene N-myc. Mol Cell Biol. 1987 May;7(5):1638–1645. doi: 10.1128/mcb.7.5.1638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  30. Stark G. R., Debatisse M., Giulotto E., Wahl G. M. Recent progress in understanding mechanisms of mammalian DNA amplification. Cell. 1989 Jun 16;57(6):901–908. doi: 10.1016/0092-8674(89)90328-0. [DOI] [PubMed] [Google Scholar]
  31. Stark G. R., Wahl G. M. Gene amplification. Annu Rev Biochem. 1984;53:447–491. doi: 10.1146/annurev.bi.53.070184.002311. [DOI] [PubMed] [Google Scholar]
  32. Tereba A., McCarthy B. J. Hybridization of 125I-labeled ribonucleic acid. Biochemistry. 1973 Nov 6;12(23):4675–4679. doi: 10.1021/bi00747a020. [DOI] [PubMed] [Google Scholar]
  33. Von Hoff D. D., Needham-VanDevanter D. R., Yucel J., Windle B. E., Wahl G. M. Amplified human MYC oncogenes localized to replicating submicroscopic circular DNA molecules. Proc Natl Acad Sci U S A. 1988 Jul;85(13):4804–4808. doi: 10.1073/pnas.85.13.4804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wahl G. M. The importance of circular DNA in mammalian gene amplification. Cancer Res. 1989 Mar 15;49(6):1333–1340. [PubMed] [Google Scholar]
  35. Wray W., Stubblefield E. A new method for the rapid isolation of chromosomes, mitotic apparatus, or nuclei from mammalian fibroblasts at near neutral pH. Exp Cell Res. 1970 Mar;59(3):469–478. doi: 10.1016/0014-4827(70)90656-7. [DOI] [PubMed] [Google Scholar]
  36. Yancopoulos G. D., Nisen P. D., Tesfaye A., Kohl N. E., Goldfarb M. P., Alt F. W. N-myc can cooperate with ras to transform normal cells in culture. Proc Natl Acad Sci U S A. 1985 Aug;82(16):5455–5459. doi: 10.1073/pnas.82.16.5455. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES