Abstract
Multiple different biologically and clinically relevant genes are often amplified in invasive breast cancer, including HER2, ESR1, CCND1, and MYC. So far, little is known about their role in tumor progression. To investigate their significance for tumor invasion, we compared pure ductal carcinoma in situ (DCIS) and DCIS associated with invasive cancer with regard to the amplification of these genes. Fluorescence in situ hybridization (FISH) was performed on a tissue microarray containing samples from 130 pure DCIS and 159 DCIS associated with invasive breast cancer. Of the latter patients, we analyzed the intraductal and invasive components separately. In addition, lymph node metastases of 23 patients with invasive carcinoma were included. Amplification rates of pure DCIS and DCIS associated with invasive cancer did not differ significantly (pure DCIS vs. DCIS associated with invasive cancer: HER2 22.7 vs. 24.2%, ESR1 19.0 vs. 24.1%, CCND1 10.0 vs. 14.8%, MYC 11.8 vs. 6.5%; P > 0.05). Furthermore, we observed a high concordance of the amplification status for all genes if in situ and invasive carcinoma of individual patients were compared. This applied also to the corresponding lymph node metastases. Our results indicate no significant differences between the gene amplification status of DCIS and invasive breast cancer concerning HER2, ESR1, CCND1, and MYC. Therefore, our data suggest an early role of all analyzed gene amplifications in breast cancer development but not in the initiation of invasive tumor growth.




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Kallioniemi OP, Kallioniemi A, Kurisu W, Thor A, Chen LC, Smith HS, Waldman FM, Pinkel D, Gray JW (1992) ERBB2 amplification in breast cancer analyzed by fluorescence in situ hybridization. Proc Natl Acad Sci USA 89:5321–5325
King CR, Kraus MH, Aaronson SA (1985) Amplification of a novel v-erbB-related gene in a human mammary carcinoma. Science 229:974–976
Berns EM, Klijn JG, van Staveren IL, Portengen H, Noordegraaf E, Foekens JA (1992) Prevalence of amplification of the oncogenes c-myc, HER2/neu, and int-2 in one thousand human breast tumours: correlation with steroid receptors. Eur J Cancer 28:697–700
Rummukainen JK, Salminen T, Lundin J, Kytola S, Joensuu H, Isola JJ (2001) Amplification of c-myc by fluorescence in situ hybridization in a population-based breast cancer tissue array. Mod Pathol 14:1030–1035
Anzick SL, Kononen J, Walker RL, Azorsa DO, Tanner MM, Guan XY, Sauter G, Kallioniemi OP, Trent JM, Meltzer PS (1997) AIB1, a steroid receptor coactivator amplified in breast and ovarian cancer. Science 277:965–968
Courjal F, Cuny M, Rodriguez C, Louason G, Speiser P, Katsaros D, Tanner MM, Zeillinger R, Theillet C (1996) DNA amplifications at 20q13 and MDM2 define distinct subsets of evolved breast and ovarian tumours. Br J Cancer 74:1984–1989
Barlund M, Forozan F, Kononen J, Bubendorf L, Chen Y, Bittner ML et al (2000) Detecting activation of ribosomal protein S6 kinase by complementary DNA and tissue microarray analysis. J Natl Cancer Inst 92:1252–1259
Luoh SW, Venkatesan N, Tripathi R (2004) Overexpression of the amplified Pip4k2beta gene from 17q11-12 in breast cancer cells confers proliferation advantage. Oncogene 23:1354–1363
Hui R, Campbell DH, Lee CS, McCaul K, Horsfall DJ, Musgrove EA, Daly RJ, Seshadri R, Sutherland RL (1997) EMS1 amplification can occur independently of CCND1 or INT-2 amplification at 11q13 and may identify different phenotypes in primary breast cancer. Oncogene 15:1617–1623
Bellacosa A, de Feo D, Godwin AK, Bell DW, Cheng JQ, Altomare DA et al (1995) Molecular alterations of the AKT2 oncogene in ovarian and breast carcinomas. Int J Cancer 64:280–285
Jarvinen TA, Tanner M, Barlund M, Borg A, Isola J (1999) Characterization of topoisomerase II alpha gene amplification and deletion in breast cancer. Genes Chromosomes Cancer 26:142–150
Bautista S, Theillet C (1998) CCND1 and FGFR1 coamplification results in the colocalization of 11q13 and 8p12 sequences in breast tumor nuclei. Genes Chromosomes Cancer 22:268–277
Ro J, North SM, Gallick GE, Hortobagyi GN, Gutterman JU, Blick M (1988) Amplified and overexpressed epidermal growth factor receptor gene in uncultured primary human breast carcinoma. Cancer Res 48:161–164
Slamon DJ, Clark GM, Wong SG, Levin WJ, Ullrich A, McGuire WL (1987) Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science 235:177–182
Sauter G, Lee J, Bartlett JM, Slamon DJ, Press MF (2009) Guidelines for human epidermal growth factor receptor 2 testing: biologic and methodologic considerations. J Clin Oncol 27:1323–1333
Wolff AC, Hammond ME, Schwartz JN, Hagerty KL, Allred DC, Cote RJ et al (2007) 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 25:118–145
Holst F, Stahl PR, Ruiz C, Hellwinkel O, Jehan Z, Wendland M et al (2007) Estrogen receptor alpha (ESR1) gene amplification is frequent in breast cancer. Nat Genet 39:655–660
Brown LA, Hoog J, Chin SF, Tao Y, Zayed AA, Chin K et al (2008) ESR1 gene amplification in breast cancer: a common phenomenon? Nat Genet 40:806–807 (author reply 810–812)
Horlings HM, Bergamaschi A, Nordgard SH, Kim YH, Han W, Noh DY et al (2008) ESR1 gene amplification in breast cancer: a common phenomenon? Nat Genet 40:807–808 (author reply 810–812)
Reis-Filho JS, Drury S, Lambros MB, Marchio C, Johnson N, Natrajan R et al (2008) ESR1 gene amplification in breast cancer: a common phenomenon? Nat Genet 40:809–810 (author reply 810–812)
Vincent-Salomon A, Raynal V, Lucchesi C, Gruel N, Delattre O (2008) ESR1 gene amplification in breast cancer: a common phenomenon? Nat Genet 40:809 (author reply 810–812)
Adelaide J, Finetti P, Charafe-Jauffret E, Wicinski J, Jacquemier J, Sotiriou C, Bertucci F, Birnbaum D, Chaffanet M (2008) Absence of ESR1 amplification in a series of breast cancers. Int J Cancer 123:2970–2972
Tomita S, Zhang Z, Nakano M, Ibusuki M, Kawazoe T, Yamamoto Y, Iwase H (2009) Estrogen receptor alpha gene ESR1 amplification may predict endocrine therapy responsiveness in breast cancer patients. Cancer Sci 100:1012–1017
Bedard PL, Piccart-Gebhart MJ (2009) Progress in tailoring adjuvant endocrine therapy for postmenopausal women with early breast cancer. Curr Opin Oncol 21:491–498
Borg A, Baldetorp B, Ferno M, Olsson H, Sigurdsson H (1992) c-myc Amplification is an independent prognostic factor in postmenopausal breast cancer. Int J Cancer 51:687–691
Champeme MH, Bieche I, Hacene K, Lidereau R (1994) Int-2/FGF3 amplification is a better independent predictor of relapse than c-myc and c-erbB-2/neu amplifications in primary human breast cancer. Mod Pathol 7:900–905
Chen Y, Olopade OI (2008) MYC in breast tumor progression. Expert Rev Anticancer Ther 8:1689–1698
Persons DL, Borelli KA, Hsu PH (1997) Quantitation of HER-2/neu and c-myc gene amplification in breast carcinoma using fluorescence in situ hybridization. Mod Pathol 10:720–727
Deming SL, Nass SJ, Dickson RB, Trock BJ (2000) C-myc amplification in breast cancer: a meta-analysis of its occurrence and prognostic relevance. Br J Cancer 83:1688–1695
Naidu R, Wahab NA, Yadav M, Kutty MK (2002) Protein expression and molecular analysis of c-myc gene in primary breast carcinomas using immunohistochemistry and differential polymerase chain reaction. Int J Mol Med 9:189–196
Berns EM, Klijn JG, van Putten WL, van Staveren IL, Portengen H, Foekens JA (1992) c-myc Amplification is a better prognostic factor than HER2/neu amplification in primary breast cancer. Cancer Res 52:1107–1113
Scorilas A, Trangas T, Yotis J, Pateras C, Talieri M (1999) Determination of c-myc amplification and overexpression in breast cancer patients: evaluation of its prognostic value against c-erbB-2, cathepsin-D and clinicopathological characteristics using univariate and multivariate analysis. Br J Cancer 81:1385–1391
Schlotter CM, Vogt U, Bosse U, Mersch B, Wassmann K (2003) C-myc, not HER-2/neu, can predict recurrence and mortality of patients with node-negative breast cancer. Breast Cancer Res 5:R30–R36
Seshadri R, Lee CS, Hui R, McCaul K, Horsfall DJ, Sutherland RL (1996) Cyclin DI amplification is not associated with reduced overall survival in primary breast cancer but may predict early relapse in patients with features of good prognosis. Clin Cancer Res 2:1177–1184
Naidu R, Wahab NA, Yadav MM, Kutty MK (2002) Expression and amplification of cyclin D1 in primary breast carcinomas: relationship with histopathological types and clinico-pathological parameters. Oncol Rep 9:409–416
Butt AJ, McNeil CM, Musgrove EA, Sutherland RL (2005) Downstream targets of growth factor and oestrogen signalling and endocrine resistance: the potential roles of c-Myc, cyclin D1 and cyclin E. Endocr Relat Cancer 12(Suppl 1):S47–S59
Robanus-Maandag EC, Bosch CA, Kristel PM, Hart AA, Faneyte IF, Nederlof PM, Peterse JL, van de Vijver MJ (2003) Association of C-MYC amplification with progression from the in situ to the invasive stage in C-MYC-amplified breast carcinomas. J Pathol 201:75–82
Watson PH, Safneck JR, Le K, Dubik D, Shiu RP (1993) Relationship of c-myc amplification to progression of breast cancer from in situ to invasive tumor and lymph node metastasis. J Natl Cancer Inst 85:902–907
Lebeau A, Unholzer A, Amann G, Kronawitter M, Bauerfeind I, Sendelhofert A, Iff A, Lohrs U (2003) EGFR, HER-2/neu, cyclin D1, p21 and p53 in correlation to cell proliferation and steroid hormone receptor status in ductal carcinoma in situ of the breast. Breast Cancer Res Treat 79:187–198
Bartkova J, Barnes DM, Millis RR, Gullick WJ (1990) Immunohistochemical demonstration of c-erbB-2 protein in mammary ductal carcinoma in situ. Hum Pathol 21:1164–1167
van de Vijver MJ, Peterse JL, Mooi WJ, Wisman P, Lomans J, Dalesio O, Nusse R (1988) Neu-protein overexpression in breast cancer. Association with comedo-type ductal carcinoma in situ and limited prognostic value in stage II breast cancer. N Engl J Med 319:1239–1245
Latta EK, Tjan S, Parkes RK, O’Malley FP (2002) The role of HER2/neu overexpression/amplification in the progression of ductal carcinoma in situ to invasive carcinoma of the breast. Mod Pathol 15:1318–1325
Soomro S, Shousha S, Taylor P, Shepard HM, Feldmann M (1991) c-erbB-2 expression in different histological types of invasive breast carcinoma. J Clin Pathol 44:211–214
Somerville JE, Clarke LA, Biggart JD (1992) c-erbB-2 overexpression and histological type of in situ and invasive breast carcinoma. J Clin Pathol 45:16–20
Allred DC, Clark GM, Molina R, Tandon AK, Schnitt SJ, Gilchrist KW, Osborne CK, Tormey DC, McGuire WL (1992) Overexpression of HER-2/neu and its relationship with other prognostic factors change during the progression of in situ to invasive breast cancer. Hum Pathol 23:974–979
Elston CW, Ellis IO (1991) Pathological prognostic factors in breast cancer. I. The value of histological grade in breast cancer: experience from a large study with long-term follow-up. Histopathology 19:403–410
WHO (2003) Tumors of the breast. In: Tavassoli FA, Devilee P (eds) World Health Organisation classification of tumors: pathology and genetics of tumours of the breast and female genital organs. IARC Press, Lyon, pp 9–112
Sauter G, Simon R, Hillan K (2003) Tissue microarrays in drug discovery. Nat Rev Drug Discov 2:962–972
Al-Kuraya K, Schraml P, Torhorst J, Tapia C, Zaharieva B, Novotny H et al (2004) Prognostic relevance of gene amplifications and coamplifications in breast cancer. Cancer Res 64:8534–8540
Kuerer HM, Albarracin CT, Yang WT, Cardiff RD, Brewster AM, Symmans WF et al (2009) Ductal carcinoma in situ: state of the science and roadmap to advance the field. J Clin Oncol 27:279–288
Glockner S, Lehmann U, Wilke N, Kleeberger W, Langer F, Kreipe H (2001) Amplification of growth regulatory genes in intraductal breast cancer is associated with higher nuclear grade but not with the progression to invasiveness. Lab Investig 81:565–571
Park K, Han S, Kim HJ, Kim J, Shin E (2006) HER2 status in pure ductal carcinoma in situ and in the intraductal and invasive components of invasive ductal carcinoma determined by fluorescence in situ hybridization and immunohistochemistry. Histopathology 48:702–707
Done SJ, Arneson NC, Ozcelik H, Redston M, Andrulis IL (1998) p53 mutations in mammary ductal carcinoma in situ but not in epithelial hyperplasias. Cancer Res 58:785–789
van der Groep P, van Diest PJ, Menko F, Bart J, de Vries E, van der Wall E (2009) Molecular profile of ductal carcinoma in situ of the breast in BRCA1 and BRCA2 germline mutation carriers. J Clin Pathol 62:926–930
Half E, Tang XM, Gwyn K, Sahin A, Wathen K, Sinicrope FA (2002) Cyclooxygenase-2 expression in human breast cancers and adjacent ductal carcinoma in situ. Cancer Res 62:1676–1681
Dietrich D, Schuster M, Lesche R, Haedicke W, Kristiansen G (2007) Multiplexed methylation analysis—a new technology to analyse the methylation pattern of laser microdissected cells of normal breast tissue, DCIS and invasive ductal carcinoma of the breast. Verh Dtsch Ges Pathol 91:197–207
Von Hoff DD, McGill JR, Forseth BJ, Davidson KK, Bradley TP, Van Devanter DR, Wahl GM (1992) Elimination of extrachromosomally amplified MYC genes from human tumor cells reduces their tumorigenicity. Proc Natl Acad Sci USA 89:8165–8169
Sontag L, Axelrod DE (2005) Evaluation of pathways for progression of heterogeneous breast tumors. J Theor Biol 232:179–189
Singletary SE (2002) A working model for the time sequence of genetic changes in breast tumorigenesis. J Am Coll Surg 194:202–216
Holst F, Stahl PR, Hellwinkel O, Dancau A, Krohn A, Wuth L et al (2008) Reply to “ESR1 gene amplification in breast cancer: a common phenomenon?”. Nat Genet 40:810–812
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This work was supported by the Deutsche Krebshilfe e. V., Germany (KH 108059).
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Burkhardt, L., Grob, T.J., Hermann, I. et al. Gene amplification in ductal carcinoma in situ of the breast. Breast Cancer Res Treat 123, 757–765 (2010). https://doi.org/10.1007/s10549-009-0675-8
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DOI: https://doi.org/10.1007/s10549-009-0675-8