Journal of Clinical Oncology, Vol 15, 2894-2904, Copyright © 1997 by American Society of Clinical Oncology

ARTICLES

HER-2/neu gene amplification characterized by fluorescence in situ hybridization: poor prognosis in node-negative breast carcinomas

MF Press, L Bernstein, PA Thomas, LF Meisner, JY Zhou, Y Ma, G Hung, RA Robinson, C Harris, A El-Naggar, DJ Slamon, RN Phillips, JS Ross, SR Wolman and KJ Flom
Norris Comprehensive Cancer Center and Department of Pathology, University of Southern California School of Medicine, Los Angeles, CA 90033, USA. villalob@hsc.usc.edu

PURPOSE: The HER-2/neu gene codes for a membrane receptor protein that is homologous, but distinct from the epidermal growth factor receptor. This investigation was performed to validate fluorescence in situ hybridization (FISH) as a sensitive and specific method for assessing HER-2/neu gene amplification in archival tissue and to test whether this alteration is associated with poor prognosis. MATERIALS AND METHODS: HER-2/neu gene amplification was determined by FISH in 140 archival breast cancers, previously characterized for gene amplification by Southern hybridization or dot-blot hybridization, and for gene expression by Northern hybridization, Western immunoblot, or immunohistochemistry. A separate cohort of 324 node-negative breast cancers was assessed for amplification by FISH to determine the utility of HER-2/neu gene amplification. RESULTS: Relative to solid-matrix blotting procedures, FISH analysis of HER-2/neu gene amplification showed a sensitivity of 98% and a specificity of 100% in 140 breast cancers. Among patients treated by surgery only, the relative risks (relative hazard) of early recurrence (recurrent disease within 24 months of diagnosis), recurrent disease (at any time), and disease- related death were statistically significantly associated with amplification. The prognostic information contributed by HER-2/neu amplification was independent of the other markers studied. CONCLUSION: FISH was an alternative technique for determining gene amplification and had some distinct advantages over Southern hybridization. Our results demonstrate that HER-2/neu gene amplification in the absence of adjuvant therapy is an independent predictor of poor clinical outcome and is a stronger discriminant than tumor size. Women with small tumors that had gene amplification were at increased risk of recurrence and disease-related death.

This article has been cited by other articles:

				J. M.S. Bartlett, M. Ibrahim, B. Jasani, J. M. Morgan, I. Ellis, E. Kay, Y. Connolly, F. Campbell, A. O'Grady, S. Barnett, et al. External Quality Assurance of HER2 FISH and ISH Testing: Three Years of the UK National External Quality Assurance Scheme Am J Clin Pathol, January 1, 2009; 131(1): 106 - 111. [Abstract] [Full Text] [PDF]

				A. Di Leo, H. L. Gomez, Z. Aziz, Z. Zvirbule, J. Bines, M. C. Arbushites, S. F. Guerrera, M. Koehler, C. Oliva, S. H. Stein, et al. Phase III, Double-Blind, Randomized Study Comparing Lapatinib Plus Paclitaxel With Placebo Plus Paclitaxel As First-Line Treatment for Metastatic Breast Cancer J. Clin. Oncol., December 1, 2008; 26(34): 5544 - 5552. [Abstract] [Full Text] [PDF]

				M. F. Press, R. S. Finn, D. Cameron, A. Di Leo, C. E. Geyer, I. E. Villalobos, A. Santiago, R. Guzman, A. Gasparyan, Y. Ma, et al. HER-2 Gene Amplification, HER-2 and Epidermal Growth Factor Receptor mRNA and Protein Expression, and Lapatinib Efficacy in Women with Metastatic Breast Cancer Clin. Cancer Res., December 1, 2008; 14(23): 7861 - 7870. [Abstract] [Full Text] [PDF]

				A. Carbone, G. Botti, A. Gloghini, G. Simone, M. Truini, M. P. Curcio, P. Gasparini, A. Mangia, T. Perin, S. Salvi, et al. Delineation of HER2 Gene Status in Breast Carcinoma by Silver in Situ Hybridization is Reproducible among Laboratories and Pathologists J. Mol. Diagn., November 1, 2008; 10(6): 527 - 536. [Abstract] [Full Text] [PDF]

				I. Vanden Bempt, P. Van Loo, M. Drijkoningen, P. Neven, A. Smeets, M.-R. Christiaens, R. Paridaens, and C. De Wolf-Peeters Polysomy 17 in Breast Cancer: Clinicopathologic Significance and Impact on HER-2 Testing J. Clin. Oncol., October 20, 2008; 26(30): 4869 - 4874. [Abstract] [Full Text] [PDF]

				S. Menard, A. Balsari, E. Tagliabue, T. Camerini, P. Casalini, R. Bufalino, F. Castiglioni, M. L. Carcangiu, A. Gloghini, S. Scalone, et al. Biology, prognosis and response to therapy of breast carcinomas according to HER2 score Ann. Onc., October 1, 2008; 19(10): 1706 - 1712. [Abstract] [Full Text] [PDF]

				M. Campos, C. Prior, F. Warleta, I. Zudaire, J. Ruiz-Mora, R. Catena, A. Calvo, and J. J. Gaforio Phenotypic and Genetic Characterization of Circulating Tumor Cells by Combining Immunomagnetic Selection and FICTION Techniques J. Histochem. Cytochem., July 1, 2008; 56(7): 667 - 675. [Abstract] [Full Text] [PDF]

				S. Kulkarni and D. G. Hicks HER2-Positive Early Breast Cancer and Trastuzumab: A Surgeon's Perspective Ann. Surg. Oncol., June 1, 2008; 15(6): 1677 - 1688. [Abstract] [Full Text] [PDF]

				K. Bullock and K. Blackwell Clinical Efficacy of Taxane-Trastuzumab Combination Regimens for HER-2-Positive Metastatic Breast Cancer Oncologist, May 1, 2008; 13(5): 515 - 525. [Abstract] [Full Text] [PDF]

				A. A. Garcia Small Molecules: Big Changes in the Cancer Treatment Paradigm Journal of Pharmacy Practice, February 1, 2008; 21(1): 17 - 35. [Abstract] [PDF]

				C. M. Croce Oncogenes and Cancer N. Engl. J. Med., January 31, 2008; 358(5): 502 - 511. [Full Text] [PDF]

				L. Harris, H. Fritsche, R. Mennel, L. Norton, P. Ravdin, S. Taube, M. R. Somerfield, D. F. Hayes, and R. C. Bast Jr American Society of Clinical Oncology 2007 Update of Recommendations for the Use of Tumor Markers in Breast Cancer J. Clin. Oncol., November 20, 2007; 25(33): 5287 - 5312. [Abstract] [Full Text] [PDF]

				J. C. Boughey, R. J. Gonzalez, E. Bonner, and H. M. Kuerer Current Treatment and Clinical Trial Developments for Ductal Carcinoma In Situ of the Breast Oncologist, November 1, 2007; 12(11): 1276 - 1287. [Abstract] [Full Text] [PDF]

				J. M S Bartlett, M. Ibrahim, K. Miller, B. Jasani, J. M Morgan, I. Ellis, E. Kay, H. Magee, and S. Barnett External quality assurance of HER2 fluorescence in situ hybridisation testing: results of a UK NEQAS pilot scheme J. Clin. Pathol., July 1, 2007; 60(7): 816 - 819. [Abstract] [Full Text] [PDF]

				K. Lehnes, A. D. Winder, C. Alfonso, N. Kasid, M. Simoneaux, H. Summe, E. Morgan, M. C. Iann, J. Duncan, M. Eagan, et al. The Effect of Estradiol on in Vivo Tumorigenesis Is Modulated by the Human Epidermal Growth Factor Receptor 2/Phosphatidylinositol 3-Kinase/Akt1 Pathway Endocrinology, March 1, 2007; 148(3): 1171 - 1180. [Abstract] [Full Text] [PDF]

				L. N. Harris, F. You, S. J. Schnitt, A. Witkiewicz, X. Lu, D. Sgroi, P. D. Ryan, S. E. Come, H. J. Burstein, B.-A. Lesnikoski, et al. Predictors of Resistance to Preoperative Trastuzumab and Vinorelbine for HER2-Positive Early Breast Cancer Clin. Cancer Res., February 15, 2007; 13(4): 1198 - 1207. [Abstract] [Full Text] [PDF]

				A. C. Wolff, M. E. H. Hammond, J. N. Schwartz, K. L. Hagerty, D. C. Allred, R. J. Cote, M. Dowsett, P. L. Fitzgibbons, W. M. Hanna, A. Langer, et al. American Society of Clinical Oncology/College of American Pathologists Guideline Recommendations for Human Epidermal Growth Factor Receptor 2 Testing in Breast Cancer J. Clin. Oncol., January 1, 2007; 25(1): 118 - 145. [Abstract] [Full Text] [PDF]

				M. D. Williams, N. Chakravarti, M. S. Kies, S.-I. Maruya, J. N. Myers, J. C. Haviland, R. S. Weber, R. Lotan, and A. K. El-Naggar Implications of Methylation Patterns of Cancer Genes in Salivary Gland Tumors Clin. Cancer Res., December 15, 2006; 12(24): 7353 - 7358. [Abstract] [Full Text] [PDF]

				A. M. Gonzalez-Angulo, G. N. Hortobagyi, and F. J. Esteva Adjuvant Therapy with Trastuzumab for HER-2/neu-Positive Breast Cancer Oncologist, September 1, 2006; 11(8): 857 - 867. [Abstract] [Full Text] [PDF]

				C. Jackisch HER-2-Positive Metastatic Breast Cancer: Optimizing Trastuzumab-Based Therapy Oncologist, September 1, 2006; 11(suppl_1): 34 - 41. [Abstract] [Full Text] [PDF]

				K. Kimura, T. Sawada, M. Komatsu, M. Inoue, K. Muguruma, T. Nishihara, Y. Yamashita, N. Yamada, M. Ohira, and K. Hirakawa Antitumor Effect of Trastuzumab for Pancreatic Cancer with High HER-2 Expression and Enhancement of Effect by Combined Therapy with Gemcitabine. Clin. Cancer Res., August 15, 2006; 12(16): 4925 - 4932. [Abstract] [Full Text] [PDF]

				N. Robert, B. Leyland-Jones, L. Asmar, R. Belt, D. Ilegbodu, D. Loesch, R. Raju, E. Valentine, R. Sayre, M. Cobleigh, et al. Randomized Phase III Study of Trastuzumab, Paclitaxel, and Carboplatin Compared With Trastuzumab and Paclitaxel in Women With HER-2-Overexpressing Metastatic Breast Cancer J. Clin. Oncol., June 20, 2006; 24(18): 2786 - 2792. [Abstract] [Full Text] [PDF]

				L. A. Carey, C. M. Perou, C. A. Livasy, L. G. Dressler, D. Cowan, K. Conway, G. Karaca, M. A. Troester, C. K. Tse, S. Edmiston, et al. Race, breast cancer subtypes, and survival in the Carolina Breast Cancer Study. JAMA, June 7, 2006; 295(21): 2492 - 2502. [Abstract] [Full Text] [PDF]

				M. Neyt, J. Albrecht, and V. Cocquyt An economic evaluation of Herceptin(R) in adjuvant setting: the Breast Cancer International Research Group 006 trial Ann. Onc., March 1, 2006; 17(3): 381 - 390. [Abstract] [Full Text] [PDF]

				H. Joensuu, P.-L. Kellokumpu-Lehtinen, P. Bono, T. Alanko, V. Kataja, R. Asola, T. Utriainen, R. Kokko, A. Hemminki, M. Tarkkanen, et al. Adjuvant docetaxel or vinorelbine with or without trastuzumab for breast cancer. N. Engl. J. Med., February 23, 2006; 354(8): 809 - 820. [Abstract] [Full Text] [PDF]

				S. Veeramani, T.-C. Yuan, S.-J. Chen, F.-F. Lin, J. E Petersen, S. Shaheduzzaman, S. Srivastava, R. G MacDonald, and M.-F. Lin Cellular prostatic acid phosphatase: a protein tyrosine phosphatase involved in androgen-independent proliferation of prostate cancer Endocr. Relat. Cancer, December 1, 2005; 12(4): 805 - 822. [Abstract] [Full Text] [PDF]

				E. Espinosa, J.A. F. Vara, A. Redondo, J.J. Sanchez, D. Hardisson, P. Zamora, F. G. Pastrana, P. Cejas, B. Martinez, A. Suarez, et al. Breast Cancer Prognosis Determined by Gene Expression Profiling: A Quantitative Reverse Transcriptase Polymerase Chain Reaction Study J. Clin. Oncol., October 10, 2005; 23(29): 7278 - 7285. [Abstract] [Full Text] [PDF]

				R Ainsworth, J M S Bartlett, J J Going, E A Mallon, A Forsyth, J Richmond, W Angerson, A Watters, and B Dunne IHC for Her2 with CBE356 antibody is a more accurate predictor of Her2 gene amplification by FISH than HercepTestTM in breast carcinoma J. Clin. Pathol., October 1, 2005; 58(10): 1086 - 1090. [Abstract] [Full Text] [PDF]

				M. F. Press, G. Sauter, L. Bernstein, I. E. Villalobos, M. Mirlacher, J.-Y. Zhou, R. Wardeh, Y.-T. Li, R. Guzman, Y. Ma, et al. Diagnostic Evaluation of HER-2 as a Molecular Target: An Assessment of Accuracy and Reproducibility of Laboratory Testing in Large, Prospective, Randomized Clinical Trials Clin. Cancer Res., September 15, 2005; 11(18): 6598 - 6607. [Abstract] [Full Text] [PDF]

				H Sartelet, E Lagonotte, M Lorenzato, I Duval, C Lechki, C Rigaud, J Cucherousset, A Durlach, O Graesslin, P Abboud, et al. Comparison of liquid based cytology and histology for the evaluation of HER-2 status using immunostaining and CISH in breast carcinoma J. Clin. Pathol., August 1, 2005; 58(8): 864 - 871. [Abstract] [Full Text] [PDF]

				J. H. Kim, V. Bossuyt, T. Ponn, D. Lannin, and B. G. Haffty Cyclooxygenase-2 Expression in Postmastectomy Chest Wall Relapse Clin. Cancer Res., July 15, 2005; 11(14): 5199 - 5205. [Abstract] [Full Text] [PDF]

				L. G. Dressler, D. A. Berry, G. Broadwater, D. Cowan, K. Cox, S. Griffin, A. Miller, J. Tse, D. Novotny, D. L. Persons, et al. Comparison of HER2 Status by Fluorescence in Situ Hybridization and Immunohistochemistry to Predict Benefit From Dose Escalation of Adjuvant Doxorubicin-Based Therapy in Node-Positive Breast Cancer Patients J. Clin. Oncol., July 1, 2005; 23(19): 4287 - 4297. [Abstract] [Full Text] [PDF]

				C. Tse, D. Brault, J. Gligorov, M. Antoine, R. Neumann, J.-P. Lotz, and J. Capeau Evaluation of the Quantitative Analytical Methods Real-Time PCR for HER-2 Gene Quantification and ELISA of Serum HER-2 Protein and Comparison with Fluorescence in Situ Hybridization and Immunohistochemistry for Determining HER-2 Status in Breast Cancer Patients Clin. Chem., July 1, 2005; 51(7): 1093 - 1101. [Abstract] [Full Text] [PDF]

				H. Huang, J. Groth, K. Sossey-Alaoui, L. Hawthorn, S. Beall, and J. Geradts Aberrant Expression of Novel and Previously Described Cell Membrane Markers in Human Breast Cancer Cell Lines and Tumors Clin. Cancer Res., June 15, 2005; 11(12): 4357 - 4364. [Abstract] [Full Text] [PDF]

				M. D. Pegram, T. Pienkowski, D. W. Northfelt, W. Eiermann, R. Patel, P. Fumoleau, E. Quan, J. Crown, D. Toppmeyer, M. Smylie, et al. Results of Two Open-Label, Multicenter Phase II Studies of Docetaxel, Platinum Salts, and Trastuzumab in HER2-Positive Advanced Breast Cancer J Natl Cancer Inst, May 19, 2004; 96(10): 759 - 769. [Abstract] [Full Text] [PDF]

				H. Burris III, D. Yardley, S. Jones, G. Houston, C. Broome, D. Thompson, F. A. Greco, M. White, and J. Hainsworth Phase II Trial of Trastuzumab Followed by Weekly Paclitaxel/Carboplatin As First-Line Treatment for Patients With Metastatic Breast Cancer J. Clin. Oncol., May 1, 2004; 22(9): 1621 - 1629. [Abstract] [Full Text] [PDF]

				H. Yaziji, L. C. Goldstein, T. S. Barry, R. Werling, H. Hwang, G. K. Ellis, J. R. Gralow, R. B. Livingston, and A. M. Gown HER-2 Testing in Breast Cancer Using Parallel Tissue-Based Methods JAMA, April 28, 2004; 291(16): 1972 - 1977. [Abstract] [Full Text] [PDF]

				J. S. Ross, J. A. Fletcher, K. J. Bloom, G. P. Linette, J. Stec, W. F. Symmans, L. Pusztai, and G. N. Hortobagyi Targeted Therapy in Breast Cancer: The HER-2/neu Gene and Protein Mol. Cell. Proteomics, April 1, 2004; 3(4): 379 - 398. [Abstract] [Full Text] [PDF]

				E. B. Elkin, M. C. Weinstein, E. P. Winer, K. M. Kuntz, S. J. Schnitt, and J. C. Weeks HER-2 Testing and Trastuzumab Therapy for Metastatic Breast Cancer: A Cost-Effectiveness Analysis J. Clin. Oncol., March 1, 2004; 22(5): 854 - 863. [Abstract] [Full Text] [PDF]

				G. E. Konecny, Y. G. Meng, M. Untch, H.-J. Wang, I. Bauerfeind, M. Epstein, P. Stieber, J.-M. Vernes, J. Gutierrez, K. Hong, et al. Association between HER-2/neu and Vascular Endothelial Growth Factor Expression Predicts Clinical Outcome in Primary Breast Cancer Patients Clin. Cancer Res., March 1, 2004; 10(5): 1706 - 1716. [Abstract] [Full Text] [PDF]

				R. Nahta and F. J. Esteva HER-2-Targeted Therapy: Lessons Learned and Future Directions Clin. Cancer Res., November 1, 2003; 9(14): 5078 - 5084. [Abstract] [Full Text] [PDF]

				J. S. Ross, J. A. Fletcher, G. P. Linette, J. Stec, E. Clark, M. Ayers, W. F. Symmans, L. Pusztai, and K. J. Bloom The HER-2/neu Gene and Protein in Breast Cancer 2003: Biomarker and Target of Therapy Oncologist, August 1, 2003; 8(4): 307 - 325. [Abstract] [Full Text] [PDF]

				A. Volpi, O. Nanni, F. De Paola, A. M. Granato, A. Mangia, F. Monti, F. Schittulli, M. De Lena, E. Scarpi, P. Rosetti, et al. HER-2 Expression and Cell Proliferation: Prognostic Markers in Patients With Node-Negative Breast Cancer J. Clin. Oncol., July 15, 2003; 21(14): 2708 - 2712. [Abstract] [Full Text] [PDF]

				L Matyakhina, S Pack, L S Kirschner, E Pak, P Mannan, J Jaikumar, S E Taymans, F Sandrini, J A Carney, and C A Stratakis Chromosome 2 (2p16) abnormalities in Carney complex tumours J. Med. Genet., April 1, 2003; 40(4): 268 - 277. [Abstract] [Full Text] [PDF]

				I. Zemzoum, R. E. Kates, J. S. Ross, P. Dettmar, M. Dutta, C. Henrichs, S. Yurdseven, H. Hofler, M. Kiechle, M. Schmitt, et al. Invasion Factors uPA/PAI-1 and HER2 Status Provide Independent and Complementary Information on Patient Outcome in Node-Negative Breast Cancer J. Clin. Oncol., March 15, 2003; 21(6): 1022 - 1028. [Abstract] [Full Text] [PDF]

				C. F. Verschraegen, W. Hu, Y. Du, J. Mendoza, J. Early, M. Deavers, R. S. Freedman, R. C. Bast Jr., A. P. Kudelka, J. J. Kavanagh, et al. Establishment and Characterization of Cancer Cell Cultures and Xenografts Derived from Primary or Metastatic Mullerian Cancers Clin. Cancer Res., February 1, 2003; 9(2): 845 - 852. [Abstract] [Full Text] [PDF]

				G. Konecny, G. Pauletti, M. Pegram, M. Untch, S. Dandekar, Z. Aguilar, C. Wilson, H.-M. Rong, I. Bauerfeind, M. Felber, et al. Quantitative Association Between HER-2/neu and Steroid Hormone Receptors in Hormone Receptor-Positive Primary Breast Cancer J Natl Cancer Inst, January 15, 2003; 95(2): 142 - 153. [Abstract] [Full Text] [PDF]

				X. Yang, S. Groshen, S. C. Formenti, N. E. Davidson, and M. F. Press P7 Antigen Expression in Human Breast Cancer Clin. Cancer Res., January 1, 2003; 9(1): 201 - 206. [Abstract] [Full Text] [PDF]

				P. Birner, G. Oberhuber, M. F. Press, L. Bernstein, K. Flom, and D. Slamon Is Fluorescence In Situ Hybridization Really Superior to HercepTest? J. Clin. Oncol., December 1, 2002; 20(23): 4607 - 4609. [Full Text] [PDF]

				M. F. Press, D. J. Slamon, K. J. Flom, J. Park, J.-Y. Zhou, and L. Bernstein Evaluation of HER-2/neu Gene Amplification and Overexpression: Comparison of Frequently Used Assay Methods in a Molecularly Characterized Cohort of Breast Cancer Specimens J. Clin. Oncol., July 15, 2002; 20(14): 3095 - 3105. [Abstract] [Full Text] [PDF]

				K. Subbaramaiah, L. Norton, W. Gerald, and A. J. Dannenberg Cyclooxygenase-2 Is Overexpressed in HER-2/neu-positive Breast Cancer. EVIDENCE FOR INVOLVEMENT OF AP-1 AND PEA3 J. Biol. Chem., May 17, 2002; 277(21): 18649 - 18657. [Abstract] [Full Text] [PDF]

				R. Tubbs, J. Pettay, M. Skacel, R. Powell, M. Stoler, P. Roche, and J. Hainfeld Gold-Facilitated in Situ Hybridization : A Bright-Field Autometallographic Alternative to Fluorescence in Situ Hybridization for Detection of HER-2/neu Gene Amplification Am. J. Pathol., May 1, 2002; 160(5): 1589 - 1595. [Abstract] [Full Text] [PDF]

				J. S. Coon, E. Marcus, S. Gupta-Burt, S. Seelig, K. Jacobson, S. Chen, V. Renta, G. Fronda, and H. D. Preisler Amplification and Overexpression of Topoisomerase II{alpha} Predict Response to Anthracycline-based Therapy in Locally Advanced Breast Cancer Clin. Cancer Res., April 1, 2002; 8(4): 1061 - 1067. [Abstract] [Full Text] [PDF]

				M. Colleoni, N. Rotmensz, C. Robertson, L. Orlando, G. Viale, G. Renne, A. Luini, P. Veronesi, M. Intra, R. Orecchia, et al. Very young women (<35 years) with operable breast cancer: features of disease at presentation Ann. Onc., February 20, 2002; 13(2): 273 - 279. [Abstract] [Full Text] [PDF]

				N. Harbeck, R. E. Kates, and M. Schmitt Clinical Relevance of Invasion Factors Urokinase-Type Plasminogen Activator and Plasminogen Activator Inhibitor Type 1 for Individualized Therapy Decisions in Primary Breast Cancer Is Greatest When Used in Combination J. Clin. Oncol., February 15, 2002; 20(4): 1000 - 1007. [Abstract] [Full Text] [PDF]