ERBB2 amplification in breast cancer analyzed by fluorescence in situ hybridization

Olli P. Kallioniemi, Anne Kallioniemi, Wayne Kurisu, Ann Thor, Ling Chun Chen, Helene S. Smith, Frederic M. Waldman, Dan Pinkel, Joe Gray

Research output: Contribution to journalArticle

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Abstract

We illustrate the use of fluorescence in situ hybridization (FISH) for analysis of ERBB2 oncogene copy number, the level of amplification (here defined as the ratio of ERBB2 copy number to copy number of chromosome 17 centromeres), and the distribution of amplified genes in breast cancer cell lines and uncultured primary breast carcinomas. The relative ERBB2 copy number determined by FISH in 10 breast cancer cell lines correlated strongly with Southern blot results (r = 0.98) when probes for an identical reference locus were used in the two methods. Metaphase analysis of cell lines showed that amplified ERBB2 copies always occurred in intrachromosomal clusters but that the number and chromosomal location of these clusters varied among the cell lines. In interphase nuclei of primary tumors showing ERBB2 amplification (10/44), ERBB2 copies were seen as one to four clusters, also suggesting intrachromosomal localization. Regardless of the average level of amplification, all these tumors contained highly amplified cell subpopulations with at least 25, and sometimes more than 100, ERBB2 copies per cell. Tumors that did not show amplification by FISH (34/44) had an average of one to five ERBB2 copies scattered randomly in the nuclei and completely lacked cells with high copy levels. FISH results on primary tumors were concordant with slot blot results on amplification and with immunohistochemical detection of over-expression. Quantitative analysis of ERBB2 amplification by FISH may improve prognostic assessments based on the pattern of amplification and detection of heavily amplified tumor cell subpopulations.

Original languageEnglish (US)
Pages (from-to)5321-5325
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume89
Issue number12
StatePublished - 1992
Externally publishedYes

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Fluorescence In Situ Hybridization
Breast Neoplasms
Cell Line
Neoplasms
Chromosomes, Human, Pair 17
Centromere
Interphase
Metaphase
Southern Blotting
Oncogenes
Genes

ASJC Scopus subject areas

  • Genetics
  • General

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ERBB2 amplification in breast cancer analyzed by fluorescence in situ hybridization. / Kallioniemi, Olli P.; Kallioniemi, Anne; Kurisu, Wayne; Thor, Ann; Chen, Ling Chun; Smith, Helene S.; Waldman, Frederic M.; Pinkel, Dan; Gray, Joe.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 89, No. 12, 1992, p. 5321-5325.

Research output: Contribution to journalArticle

Kallioniemi, OP, Kallioniemi, A, Kurisu, W, Thor, A, Chen, LC, Smith, HS, Waldman, FM, Pinkel, D & Gray, J 1992, 'ERBB2 amplification in breast cancer analyzed by fluorescence in situ hybridization', Proceedings of the National Academy of Sciences of the United States of America, vol. 89, no. 12, pp. 5321-5325.
Kallioniemi, Olli P. ; Kallioniemi, Anne ; Kurisu, Wayne ; Thor, Ann ; Chen, Ling Chun ; Smith, Helene S. ; Waldman, Frederic M. ; Pinkel, Dan ; Gray, Joe. / ERBB2 amplification in breast cancer analyzed by fluorescence in situ hybridization. In: Proceedings of the National Academy of Sciences of the United States of America. 1992 ; Vol. 89, No. 12. pp. 5321-5325.
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AU - Kurisu, Wayne

AU - Thor, Ann

AU - Chen, Ling Chun

AU - Smith, Helene S.

AU - Waldman, Frederic M.

AU - Pinkel, Dan

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