title: Prognostic value of the molecular detection of circulating tumor cells using a multimarker reverse transcription-PCR assay for cytokeratin 19, mammaglobin A, and HER2 in early breast cancer.
abstract: OBJECTIVE
  To investigate the prognostic value of the molecular detection of circulating tumor cells (CTCs) using three markers [cytokeratin 19 (CK19), mammaglobin A (MGB1), and HER2] in early breast cancer.
  
  
  METHODS
  CK19mRNA+, MGB1mRNA+, and HER2mRNA+ cells were detected using real-time (CK19) and nested (MGB1 and HER2) reverse transcription-PCR in the peripheral blood of 175 women with stage I to III breast cancer before the initiation of adjuvant chemotherapy. The detection of CTCs was correlated with clinical outcome. In 10 patients, immunofluorescence staining experiments were done to investigate the coexpression of cytokeratin, MGB1, and HER2 in CTCs.
  
  
  RESULTS
  CK19mRNA+, MGB1mRNA+, and HER2mRNA+ cells were detected in 41.1%, 8%, and 28.6% of the 175 patients, respectively. Patients had one of the following molecular profiles: CK19mRNA+/MGB1mRNA+/HER2mRNA+ (n = 8), CK19mRNA+/MGB1mRNA+/HER2mRNA- (n = 1), CK19mRNA+/MGB1mRNA-/HER2mRNA+ (n = 42), CK19mRNA+/MGB1mRNA-/HER2mRNA- (n = 21), CK19mRNA-/MGB1mRNA+/HER2mRNA- (n = 5), and CK19mRNA-/MGB1mRNA-/HER2mRNA- (n = 98). Double-immunofluorescence experiments confirmed the following CTC phenotypes: CK+/MGB1+, CK+/MGB1-, CK-/MGB1+, CK+/HER2+, CK+/HER2-, MGB1+/HER2-, and MGB1+/HER2+. In univariate analysis, the detection of CK19mRNA+, MGB1mRNA+, and HER2mRNA+ cells was associated with shorter disease-free survival (DFS; P < 0.001, P = 0.001, and P < 0.001, respectively), whereas the detection of CK19mRNA+ and MGB1mRNA+ cells was associated with worse overall survival (P = 0.044 and 0.034, respectively). In multivariate analysis, estrogen receptor-negative tumors and the detection of CK19mRNA+ and MGB1mRNA+ cells were independently associated with worse DFS.
  
  
  CONCLUSIONS
  The detection of peripheral blood CK19mRNA+ and MGB1mRNA+ cells before adjuvant chemotherapy predicts poor DFS in women with early breast cancer.
authors.affiliation: Department of Medical Oncology, University General Hospital of Heraklion, Heraklion, Crete, Greece. mavrudis@med.uoc.gr
authors.forename: Michail
authors.initials: M
authors.lastname: Ignatiadis
authors.forename: Galatea
authors.initials: G
authors.lastname: Kallergi
authors.forename: Maria
authors.initials: M
authors.lastname: Ntoulia
authors.forename: Maria
authors.initials: M
authors.lastname: Perraki
authors.forename: Stella
authors.initials: S
authors.lastname: Apostolaki
authors.forename: Maria
authors.initials: M
authors.lastname: Kafousi
authors.forename: Grigorios
authors.initials: G
authors.lastname: Chlouverakis
authors.forename: Efstathios
authors.initials: E
authors.lastname: Stathopoulos
authors.forename: Evi
authors.initials: E
authors.lastname: Lianidou
authors.forename: Vassilis
authors.initials: V
authors.lastname: Georgoulias
authors.forename: Dimitris
authors.initials: D
authors.lastname: Mavroudis
mesh_terms: D000328:Adult; D000368:Aged; D014408:Biomarkers, Tumor / Q000032:analysis; D001943:Breast Neoplasms / Q000097:blood / Q000737:chemistry / Q000175:diagnosis* / Q000473:pathology; D018450:Disease Progression; D005260:Female; D006801:Humans; D053539:Keratin-19 / Q000032:analysis* / Q000097:blood; D060785:Mammaglobin A; D008875:Middle Aged; D009363:Neoplasm Proteins / Q000032:analysis* / Q000097:blood; D009360:Neoplastic Cells, Circulating; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000032:analysis* / Q000097:blood; D014598:Uteroglobin / Q000032:analysis* / Q000097:blood
chemical_list: D014408:Biomarkers, Tumor; D053539:Keratin-19; D060785:Mammaglobin A; D009363:Neoplasm Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases; D014598:Uteroglobin; C556448:SCGB2A2 protein, human