Background: The polycomb group protein enhancer of zeste homologue 2 (EZH 2) has been reported as a marker of aggressive breast cancer. The aim of this study was to investigate the correlation between the expression of EZH 2 with p53 and Ki-67 expression and other clinicopathological parameters in primary breast carcinomas in order to determine the role of the above marker as a prognosticator of tumor aggressiveness and patient outcome. Patients and Methods: One hundred primary operable breast cancer patients were investigated in order to identify the expression of EZH 2, Ki-67 and p53 in imprint smears immunocytochemically. The prevalence of expression of these markers was then correlated with clinicopathological parameters. Follow-up was available for all patients. Results: EZH 2 was expressed in 64% of the cases and correlated with higher levels of p53 (relative risk = 3.00, p < 0.0001) and Ki-67 (relative risk = 3.25, p < 0.0001). Malignant cells showed immunoreactivity for all markers in the nucleus. Univariate analysis revealed a strong association between EZH 2 protein expression and tumor grade and size, lymph node metastasis, and HER-2 and estrogen and progesterone receptor status. Multivariable statistical analysis revealed that lymph node metastasis was the main predictor for EZH 2 expression. Decreased patient survival was also significantly associated with EZH 2 expression (p < 0.0001). Conclusions: EZH 2 expression may be a marker of poor prognosis in breast carcinoma patients and has been suggested as a candidate for targeted therapy.

1.
Daidone MG, Povradiso A, Gion M, Harbeck N, Sweep F, Schmitt M: Biomolecular features of clinical relevance in breast cancer. Eur J Nucl Med Mol Imaging 2004;31(suppl 1):S13–S14.
2.
Mitra AP, Lin H, Cote RJ, Datar RH: Biomarker profiling for cancer diagnosis, prognosis and therapeutic management. Natl Med J India 2005;18:304–312.
3.
Krajewski S, Krajewska M, Turner BC, Pratt C, Howard B, Zapata JM, Frenkel V, Robertson S, Ionov Y, Yamamoto H, Povrucho M, Takayama S, Reed JC: Prognostic significance of apoptosis regulators in breast cancer. Endocr Relat Cancer 1999;6:29–40.
4.
Madjd Z, Mehrjerdi AZ, Sharifi AM, Molanaei S, Shahzadi SZ, Asadi-Lari M: CD44+ cancer cells express higher levels of the antiapoptotic protein Bcl-2 in breast tumors. Cancer Immun 2009;9:4.
5.
Tan PH, Bay BH, Yip G, Selvovrajan S, Tan P, Wu J, Lee CH, Li KB: Immunohistochemical detection of Ki67 in breast cancer correlates with transcriptional regulation of genes related to apoptosis and cell death. Mod Pathol 2005;18:374–381.
6.
Daidone MG, Luisi A, Veneroni S, Benini E, Silvestrini R: Clinical studies of Bcl 2 and treatment benefit in breast cancer patients. Endocr Relat Cancer 1999;6:61–68.
7.
Dalquen P, Baschiera B, Chaffard R, Dietrich H, Feichter GE, Kremer K, Torhosst J: MIB-1(Ki-67) immunostaining of breast cancer cells in cytologic smears. Acta Cytol 1997;41:229–237.
8.
Itaya M, Yoshimoto J, Kojima K, Kawasaki S: Immunohistochemistry of p53, Bcl-2 and Ki-67 as predictors of chemosensitivity. Methods Mol Med 2005;110:213–227.
9.
Sherr CJ, McCormick F: The RB and p53 pathways in cancer. Cancer Cell 2002;2:103–112.
10.
Zeidler M, Kleer C: The Polycomb group protein Enhancer of Zeste 2: its links to DNA repair and breast cancer. J Mol Histol 2006;37:219–223.
11.
Tonini T, D’andrilli G, Fucito A, Gaspah L, Bagella L: Importance of EZH 2 polycomp protein in tumorigenesis process interfering with the pathway of growth suppressive key elements. J Cell Physiol 2008;214:295–300.
12.
Ding L, Kleer C: Enhancer of Zeste 2 as a marker of preneoplastic progression in the breast. Cancer Res 2006;66:9352–9355.
13.
Wei Y, Xia W, Zhang Z, Liu J, Wanh H, Adsay N, Abbarracin C, Yu D, Abbruzzese J, Mills G, Bast R, Hortobagyu G, Hung MC: Loss of trimethylation at lysine 27 of histone H3 is a predictor of poor outcome in breast, ovarian, and pancreatic cancers. Mol Carcinog 2008;47:701–706.
14.
Simon JA, Lange CA: Roles of the EZH2 histone methyltransferase in cancer epigenetics. Mutat Res 2008;647:21–29.
15.
Koss LG, Melamed R (eds): Koss’ Diagnostic Cytology and Its Histopathologic Bases, ed 5. Philadelphia, Lippincott Williams &amp; Wilkins, 2005, vol 2, pp 1592–1593, 1615–1616.
16.
Tavassoli F, Devilee P (eds): World Health Organization Classification of Tumours: Pathology and Genetics of Tumours of the Breast and Female Genital Organs. Lyon, IARC Press, 2003, p 10.
17.
Page DL, Anderson TJ: Diagnostic Histopathology of the Breast. Edinburgh, Churchill Livingstone, 1987, pp 303–307.
18.
Cytology Subgroup of the National Coordinating Committee for Breast Cancer Screening Pathology: Guidelines for cytology procedures and reporting on fine needle aspirates of the breast. Cytopathology 1994;5:316–334.
19.
Bast RC, Raudin P, Hayes DF, et al: 2000 update of recommendations for the use of tumor markers in breast and colorectal cancer: clinical practice guidelines of the American Society of Clinical Oncology. J Clin Oncol 2001;19:1865–1878.
20.
Gonzalez-Angulo AM, Morales-Vasquez F, Hortobagyi GN: Overview of resistance to systemic therapy in patients with breast cancer. Adv Exp Med Biol 2007;608:1–22.
21.
Tsakonas G, Kosmas C: Integration of novel targeted therapies into the systemic treatment of breast cancer – a review. J BUON 2007;12:319–327.
22.
Cotter TG: Apoptosis and cancer: the genesis of a research field. Nat Rev Cancer 2009;9:501–507.
23.
Shen Y, White E: p53 dependent apoptosis pathways. Adv Cancer Res 2001;82:55–84.
24.
Wiesner F, Magener A, Fasching P, Wesse J, Bani M, Rauh C, Jud S, Schraceder M, Loehberg C, Beckmann M, Hartmann A, Lux M: Ki-67 as a prognostic molecular marker in routine clinical use in breast cancer patients. Breast 2009;18:135–141.
25.
von Minckwitz G, Sinn HP, Raab G, Loibl S, Blohmer JU, Eidtmann H, Hilfrich J, Merkle E, Jackisch C, Costa S, Caputo A, Kaufmann M, German Breast Group: Clinical response after two cycles compared to HER2, Ki-67, p53, and bcl-2 in independently predicting a pathological complete response after preoperative chemotherapy in patients with operable carcinoma of the breast. Breast Cancer Res 2008;10:R30.
26.
Jacquemier J, Charafe-Jauffret E, Monville F, Esterni B, Extra J, Houvenaeghel G, Xerri L, Bertucci F, Birnbaum D: Association of GATA3, p53, Ki67 status and vascular peritumoral invasion are strongly prognostic in luminal breast cancer. Breast Cancer Res 2009;11:R23.
27.
Collett K, Eide G, Ames H, Stefansson I, Eide J, Brocaten A, Aas T, Otte A, Akslen L: Expression of Enhancer of Zeste homologue 2 is significantly associated with increased tumor cell proliferation and is a marker of aggressive breast cancer. Clin Cancer Res 2006;12:1168–1175.
28.
Mattiou E, Vogiatzi P, Sun A, Abbadessa G, Angeloni G, D’ugo D, Trani D, Gaughan J, Vecchio F, Cevenini G, Persiani R, Giordano A, Claudio P: Immunohistochemical analysis of pRb2/p130, VEGF, EZH 2, p53, p16INK4A, p27KIP1, p21WAF1, Ki-67 expression patterns in gastric cancer. J Cell Physiol 2007;210:183–191.
29.
Takeshita F, Minakuchi Y, Naghahara S, Honma K, Sasaki H, Hirai K, Teratani T, Namatame N, Yamamoto Y, Hanai K, Kato T, Sano A, Ochiya T: Efficient delivery of small interfering RNA to bone-metastatic tumors by using atelocollagen in vivo. Proc Natl Acad Sci USA 2005;102:12177–12182.
30.
Machmann I, Haluorsen O, Collett K, Stefansson I, Straume O, Haukaas S, Salvesen H, Otte A, Akslen L: EZH 2 expression is associated with high proliferation rate and aggressive tumor subgroups in cutaneous melanoma and cancers of the endometrium, prostate and breast. J Clin Oncol 2006;24:268–273.
31.
Kleer C, Cao Q, Varambally S, Shen R, Ota I, Tomlins S, Ghosh D, Sewalt R, Otte A, Hayes D, Sabel M, Livant A, Weiss S, Rubin M, Chinnaiyan A: EZH2 is a marker of aggressive breast cancer and promotes neoplastic transformation of breast epithelial cells. Proc Natl Acad Sci USA 2003;100:11606–11611.
32.
Raaphorst FM, Meijer CJ, Fieret E, et al: Poorly differentiated breast carcinoma is associated with increased expression of the human polycomb group EZH 2 gene. Neoplasia 2003;5:481–488.
33.
van’t Veer LJ, Dai H, van de Vijver M, He Y, Hart A, Mao M, Peterse H, van der Kooy K, Marton M, Witteveen A, et al: Gene expression profiling predicts clinical outcome of breast cancer. Nature 2002;415:530–536.
34.
van der Vlag J, Otte A: Transcriptional repression mediated by the human polycomb-group protein EED involves histone deacetylation. Nat Genet 1999;23:474–478.
35.
Beier R, Burgin A, Kiermaier A, et al: Induction of cyclin E-cdK2 kinase activity, E2F-dependent transcription and cell growth by Myc are genetically separable events. EMBO J 2000;19:5813–5823.
36.
Bracken AP, Pasini D, Capra M, et al: EZH 2 is downstream of the pRB-E2F pathway, essential for proliferation and amplified in cancer. EMBO J 2003;22:5323–5335.
37.
Tang X, Milyavsky M, Shats I, et al: Activated p53 suppresses the histone methyltransferase EZH 2 gene. Oncogene 2004;23:5759–5769.
38.
Norberg T, Klaar S, Kourf G, et al: Increased p53 mutation frequency during tumor progression-results from a breast cancer cohort. Cancer Res 2001;61:8317–8321.
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