title: Decreased expression of angiogenesis antagonist EFEMP1 in sporadic breast cancer is caused by aberrant promoter methylation and points to an impact of EFEMP1 as molecular biomarker.
abstract: EGF-containing fibulin-like extracellular matrix protein 1 (EFEMP1) was recently described as an antagonist of angiogenesis. Motivated by a strong dependence of tumor growth and metastasis on angiogenesis, we investigated the role of EFEMP1 in human breast cancer. We applied RNA microarray expression analysis and quantitative real-time PCR (QRT) in a total of 45 sporadic breast cancer tissues and found EFEMP1 down-regulation in 59% and 61% of the analyzed tissues, respectively. This down-regulation was confirmed on protein level. Immunohistochemistry in 211 breast cancer tissues resulted in reduced or even abolished EFEMP1 expression in 57-62.5% of the tumors. Bisulphite genomic sequencing in breast cancer cell lines and primary breast cancer tissues revealed promoter methylation as the major cause of this down-regulation. Furthermore, analysis of 203 clinically well characterized primary breast cancers displayed a significant correlation of reduced EFEMP1 protein expression with poor disease-free (p = 0.037) and overall survival (p = 0.032), particularly in those node-positive patients who received adjuvant anthracycline-based chemotherapy, but not in those treated by either cyclophosphamide-methotrexate-5-fluorouracil (CMF) or Tamoxifen. In summary, the presented data demonstrate for the first time the reduced EFEMP1 expression on RNA and protein level in a substantial number of sporadic breast carcinomas and its correlation with epigenetic alterations. Furthermore, these data point towards a possible predictive impact of EFEMP1 expression in primary breast cancer.
authors.affiliation: Department of Obstetrics and Gynecology, Technische Universitaet Muenchen, Munich, Germany.
authors.forename: Ariane
authors.initials: A
authors.lastname: Sadr-Nabavi
authors.forename: Juliane
authors.initials: J
authors.lastname: Ramser
authors.forename: Juliane
authors.initials: J
authors.lastname: Volkmann
authors.forename: Joerg
authors.initials: J
authors.lastname: Naehrig
authors.forename: Frank
authors.initials: F
authors.lastname: Wiesmann
authors.forename: Beate
authors.initials: B
authors.lastname: Betz
authors.forename: Heide
authors.initials: H
authors.lastname: Hellebrand
authors.forename: Stefanie
authors.initials: S
authors.lastname: Engert
authors.forename: Susanne
authors.initials: S
authors.lastname: Seitz
authors.forename: Rene
authors.initials: R
authors.lastname: Kreutzfeld
authors.forename: Takako
authors.initials: T
authors.lastname: Sasaki
authors.forename: Norbert
authors.initials: N
authors.lastname: Arnold
authors.forename: Rita
authors.initials: R
authors.lastname: Schmutzler
authors.forename: Marion
authors.initials: M
authors.lastname: Kiechle
authors.forename: Dieter
authors.initials: D
authors.lastname: Niederacher
authors.forename: Nadia
authors.initials: N
authors.lastname: Harbeck
authors.forename: Edgar
authors.initials: E
authors.lastname: Dahl
authors.forename: Alfons
authors.initials: A
authors.lastname: Meindl
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D020533:Angiogenesis Inhibitors / Q000627:therapeutic use; D018931:Antineoplastic Agents, Hormonal / Q000627:therapeutic use; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000627:therapeutic use; D014408:Biomarkers, Tumor / Q000235:genetics*; D001943:Breast Neoplasms / Q000188:drug therapy / Q000235:genetics* / Q000378:metabolism; D019175:DNA Methylation / Q000235:genetics*; D004252:DNA Mutational Analysis; D044127:Epigenesis, Genetic; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis*; D005260:Female; D015870:Gene Expression; D006801:Humans; D007150:Immunohistochemistry; D019656:Loss of Heterozygosity; D008875:Middle Aged; D009389:Neovascularization, Pathologic / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D011401:Promoter Regions, Genetic / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D046888:Tissue Array Analysis
chemical_list: D020533:Angiogenesis Inhibitors; D018931:Antineoplastic Agents, Hormonal; D014408:Biomarkers, Tumor; C119630:EFEMP1 protein, human; D016326:Extracellular Matrix Proteins