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PLOS ONE  2013 

Spectrum of EGFR Gene Copy Number Changes and KRAS Gene Mutation Status in Korean Triple Negative Breast Cancer Patients

DOI: 10.1371/journal.pone.0079014

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Abstract:

Anti-epidermal growth factor receptor (EGFR) therapy has been tried in triple negative breast cancer (TNBC) patients without evaluation of molecular and clinical predictors in several randomized clinical studies. Only fewer than 20% of metastatic TNBCs showed response to anti-EGFR therapy. In order to increase the overall response rate, first step would be to classify TNBC into good or poor responders according to oncogenic mutation profiles. This study provides the molecular characteristics of TNBCs including EGFR gene copy number changes and mutation status of EGFR and KRAS gene in Korean TNBC patients. Mutation analysis for EGFR, KRAS, BRAF and TP53 from a total of 105 TNBC tissue samples was performed by direct sequencing, peptide nucleic acid-mediated PCR clamping method and real-time PCR. Copy number changes of EGFR gene were evaluated using multiplex ligation-dependent probe amplification. Out of all 105 TNBCs, 15.2% (16/105) showed EGFR copy number changes. Among them, increased or decreased EGFR copy number was detected in 13 (5 single copy gain, 2 amplification and 4 high-copy number amplification) and 3 cases (3 hemizygous deletion), respectively. The mutation frequencies of KRAS, EGFR and TP53 gene were 1.9% (G12V and G12D), 1.0% (exon 19 del) and 31.4%, respectively. There was no BRAF V600E mutation found. Future studies are needed to evaluate the clinical outcomes of TNBC patients who undergo anti-EGFR therapy according to the genetic status of EGFR.

References

[1]  Reis-Filho JS, Tutt AN (2008) Triple negative tumours: a critical review. Histopathology 52: 108–118.
[2]  Huo D, Ikpatt F, Khramtsov A, Dangou JM, Nanda R, et al. (2009) Population differences in breast cancer: survey in indigenous African women reveals over-representation of triple-negative breast cancer. J Clin Oncol 27: 4515–4521.
[3]  Kim MJ, Ro JY, Ahn SH, Kim HH, Kim SB, et al. (2006) Clinicopathologic significance of the basal-like subtype of breast cancer: a comparison with hormone receptor and Her2/neu-overexpressing phenotypes. Hum Pathol 37: 1217–1226.
[4]  Kurebayashi J, Moriya T, Ishida T, Hirakawa H, Kurosumi M, et al. (2007) The prevalence of intrinsic subtypes and prognosis in breast cancer patients of different races. Breast 16 Suppl 2S72–77.
[5]  Stead LA, Lash TL, Sobieraj JE, Chi DD, Westrup JL, et al. (2009) Triple-negative breast cancers are increased in black women regardless of age or body mass index. Breast Cancer Res 11: R18.
[6]  Yin WJ, Lu JS, Di GH, Lin YP, Zhou LH, et al. (2009) Clinicopathological features of the triple-negative tumors in Chinese breast cancer patients. Breast Cancer Res Treat 115: 325–333.
[7]  Carey L, Winer E, Viale G, Cameron D, Gianni L (2010) Triple-negative breast cancer: disease entity or title of convenience? Nat Rev Clin Oncol 7: 683–692.
[8]  Hudis CA, Gianni L (2011) Triple-negative breast cancer: an unmet medical need. Oncologist 16 Suppl 11–11.
[9]  Fukui T, Mitsudomi T (2008) Mutations in the epidermal growth factor receptor gene and effects of EGFR-tyrosine kinase inhibitors on lung cancers. Gen Thorac Cardiovasc Surg 56: 97–103.
[10]  Sequist LV, Bell DW, Lynch TJ, Haber DA (2007) Molecular predictors of response to epidermal growth factor receptor antagonists in non-small-cell lung cancer. J Clin Oncol 25: 587–595.
[11]  Paez JG, Janne PA, Lee JC, Tracy S, Greulich H, et al. (2004) EGFR mutations in lung cancer: correlation with clinical response to gefitinib therapy. Science 304: 1497–1500.
[12]  Lynch TJ, Bell DW, Sordella R, Gurubhagavatula S, Okimoto RA, et al. (2004) Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib. N Engl J Med 350: 2129–2139.
[13]  Lv S, Teugels E, Sadones J, De Brakeleer S, Duerinck J, et al.. (2012) Correlation of EGFR, IDH1 and PTEN status with the outcome of patients with recurrent glioblastoma treated in a phase II clinical trial with the EGFR-blocking monoclonal antibody cetuximab. Int J Oncol.
[14]  Araki T, Shimizu K, Nakamura T, Baba M, Kawai Y, et al. (2011) Clinical screening assay for EGFR exon 19 mutations using PNA-clamp smart amplification process version 2 in lung adenocarcinoma. Oncol Rep 26: 1213–1219.
[15]  Grob TJ, Heilenkotter U, Geist S, Paluchowski P, Wilke C, et al. (2012) Rare oncogenic mutations of predictive markers for targeted therapy in triple-negative breast cancer. Breast Cancer Res Treat 134: 561–567.
[16]  Sanchez-Munoz A, Gallego E, de Luque V, Perez-Rivas LG, Vicioso L, et al. (2010) Lack of evidence for KRAS oncogenic mutations in triple-negative breast cancer. BMC Cancer 10: 136.
[17]  Jacot W, Lopez-Crapez E, Thezenas S, Senal R, Fina F, et al. (2011) Lack of EGFR-activating mutations in European patients with triple-negative breast cancer could emphasise geographic and ethnic variations in breast cancer mutation profiles. Breast Cancer Res 13: R133.
[18]  Teng YH, Tan WJ, Thike AA, Cheok PY, Tse GM, et al. (2011) Mutations in the epidermal growth factor receptor (EGFR) gene in triple negative breast cancer: possible implications for targeted therapy. Breast Cancer Res 13: R35.
[19]  Toyama T, Yamashita H, Kondo N, Okuda K, Takahashi S, et al. (2008) Frequently increased epidermal growth factor receptor (EGFR) copy numbers and decreased BRCA1 mRNA expression in Japanese triple-negative breast cancers. BMC Cancer 8: 309.
[20]  Shah SP, Roth A, Goya R, Oloumi A, Ha G, et al. (2012) The clonal and mutational evolution spectrum of primary triple-negative breast cancers. Nature 486: 395–399.
[21]  Harvey JM, Clark GM, Osborne CK, Allred DC (1999) Estrogen receptor status by immunohistochemistry is superior to the ligand-binding assay for predicting response to adjuvant endocrine therapy in breast cancer. J Clin Oncol 17: 1474–1481.
[22]  Mitsudomi T, Yatabe Y (2010) Epidermal growth factor receptor in relation to tumor development: EGFR gene and cancer. FEBS J 277: 301–308.
[23]  Olivier M, Langerod A, Carrieri P, Bergh J, Klaar S, et al. (2006) The clinical value of somatic TP53 gene mutations in 1,794 patients with breast cancer. Clin Cancer Res 12: 1157–1167.
[24]  Olivier M, Hainaut P (2001) TP53 mutation patterns in breast cancers: searching for clues of environmental carcinogenesis. Semin Cancer Biol 11: 353–360.
[25]  Normanno N, Tejpar S, Morgillo F, De Luca A, Van Cutsem E, et al. (2009) Implications for KRAS status and EGFR-targeted therapies in metastatic CRC. Nat Rev Clin Oncol 6: 519–527.
[26]  Langer CJ (2011) Roles of EGFR and KRAS Mutations in the Treatment Of Patients With Non-Small-Cell Lung Cancer. P T 36: 263–279.
[27]  Di Fiore F, Blanchard F, Charbonnier F, Le Pessot F, Lamy A, et al. (2007) Clinical relevance of KRAS mutation detection in metastatic colorectal cancer treated by Cetuximab plus chemotherapy. Br J Cancer 96: 1166–1169.
[28]  Kwon MJ, Lee SE, Kang SY, Choi YL (2011) Frequency of KRAS, BRAF, and PIK3CA mutations in advanced colorectal cancers: Comparison of peptide nucleic acid-mediated PCR clamping and direct sequencing in formalin-fixed, paraffin-embedded tissue. Pathol Res Pract 207: 762–768.
[29]  Han HS, Lim SN, An JY, Lee KM, Choe KH, et al. (2012) Detection of EGFR mutation status in lung adenocarcinoma specimens with different proportions of tumor cells using two methods of differential sensitivity. J Thorac Oncol 7: 355–364.
[30]  Wang J, Ramakrishnan R, Tang Z, Fan W, Kluge A, et al. (2010) Quantifying EGFR alterations in the lung cancer genome with nanofluidic digital PCR arrays. Clin Chem 56: 623–632.
[31]  Helfrich BA, Raben D, Varella-Garcia M, Gustafson D, Chan DC, et al. (2006) Antitumor activity of the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor gefitinib (ZD1839, Iressa) in non-small cell lung cancer cell lines correlates with gene copy number and EGFR mutations but not EGFR protein levels. Clin Cancer Res 12: 7117–7125.
[32]  Vargas AC, Reed AE, Waddell N, Lane A, Reid LE, et al.. (2012) Gene expression profiling of tumour epithelial and stromal compartments during breast cancer progression. Breast Cancer Res Treat.
[33]  Jeuken J, Cornelissen S, Boots-Sprenger S, Gijsen S, Wesseling P (2006) Multiplex ligation-dependent probe amplification: a diagnostic tool for simultaneous identification of different genetic markers in glial tumors. J Mol Diagn 8: 433–443.
[34]  Jeuken J, Sijben A, Alenda C, Rijntjes J, Dekkers M, et al. (2009) Robust detection of EGFR copy number changes and EGFR variant III: technical aspects and relevance for glioma diagnostics. Brain Pathol 19: 661–671.
[35]  Cho Y, Gorina S, Jeffrey PD, Pavletich NP (1994) Crystal structure of a p53 tumor suppressor-DNA complex: understanding tumorigenic mutations. Science 265: 346–355.
[36]  Nakajima H, Ishikawa Y, Furuya M, Sano T, Ohno Y, et al.. (2012) Protein expression, gene amplification, and mutational analysis of EGFR in triple-negative breast cancer. Breast Cancer.
[37]  Lievre A, Bachet JB, Le Corre D, Boige V, Landi B, et al. (2006) KRAS mutation status is predictive of response to cetuximab therapy in colorectal cancer. Cancer Res 66: 3992–3995.
[38]  Sok JC, Coppelli FM, Thomas SM, Lango MN, Xi S, et al. (2006) Mutant epidermal growth factor receptor (EGFRvIII) contributes to head and neck cancer growth and resistance to EGFR targeting. Clin Cancer Res 12: 5064–5073.
[39]  Moscatello DK, Holgado-Madruga M, Godwin AK, Ramirez G, Gunn G, et al. (1995) Frequent expression of a mutant epidermal growth factor receptor in multiple human tumors. Cancer Res 55: 5536–5539.
[40]  Tang CK, Gong XQ, Moscatello DK, Wong AJ, Lippman ME (2000) Epidermal growth factor receptor vIII enhances tumorigenicity in human breast cancer. Cancer Res 60: 3081–3087.
[41]  Di Fiore F, Sesboue R, Michel P, Sabourin JC, Frebourg T (2010) Molecular determinants of anti-EGFR sensitivity and resistance in metastatic colorectal cancer. Br J Cancer 103: 1765–1772.
[42]  Oden-Gangloff A, Di Fiore F, Bibeau F, Lamy A, Bougeard G, et al. (2009) TP53 mutations predict disease control in metastatic colorectal cancer treated with cetuximab-based chemotherapy. Br J Cancer 100: 1330–1335.
[43]  Powell B, Soong R, Iacopetta B, Seshadri R, Smith DR (2000) Prognostic significance of mutations to different structural and functional regions of the p53 gene in breast cancer. Clin Cancer Res 6: 443–451.
[44]  Rakha EA, El-Sayed ME, Green AR, Lee AH, Robertson JF, et al. (2007) Prognostic markers in triple-negative breast cancer. Cancer 109: 25–32.
[45]  Dickler MN, Cobleigh MA, Miller KD, Klein PM, Winer EP (2009) Efficacy and safety of erlotinib in patients with locally advanced or metastatic breast cancer. Breast Cancer Res Treat 115: 115–121.
[46]  Carey LA, Rugo HS, Marcom PK, Mayer EL, Esteva FJ, et al.. (2012) TBCRC 001: Randomized Phase II Study of Cetuximab in Combination With Carboplatin in Stage IV Triple-Negative Breast Cancer. J Clin Oncol.
[47]  Brand TM, Iida M, Wheeler DL (2011) Molecular mechanisms of resistance to the EGFR monoclonal antibody cetuximab. Cancer Biol Ther 11: 777–792.
[48]  Moran T, Sequist LV (2012) Timing of Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor Therapy in Patients With Lung Cancer With EGFR Mutations. J Clin Oncol.

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