title: Detection of Mammaglobin A-mRNA-positive circulating tumor cells in peripheral blood of patients with operable breast cancer with nested RT-PCR.
abstract: OBJECTIVE
  The development and validation of a nested RT-PCR methodology for the detection of Mammaglobin A-mRNA-positive circulating tumor cells in peripheral blood of patients with operable breast cancer and evaluation of its prognostic significance.
  
  
  METHODS
  Different combinations of specific primers were in silico designed and selected, so that false positive results due to genomic DNA contamination were avoided. The specificity of the primers used was evaluated in 30 healthy individuals, 20 patients with colorectal cancer and 20 patients with non-small cell lung cancer. The method was applied in 101 patients with operable breast cancer before the administration of adjuvant chemotherapy and 39 patients with metastatic breast cancer.
  
  
  RESULTS
  Mammaglobin A-mRNA-positive cells were detected in 14/101 (13.9%) of early breast cancer patients but not in the control population studied (0%); 9 of them (64.3%) relapsed during the follow-up period. Mammaglobin A was detected in 7/39 (17.9%) of patients with verified metastasis. Multivariate analysis revealed the detection of Mammaglobin A-mRNA-positive cells, as an independent risk factor for reduced DFI.
  
  
  CONCLUSIONS
  Mammaglobin A is a highly specific molecular marker for the detection of circulating tumor cells in operable breast cancer, with important prognostic applications.
authors.affiliation: Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, Greece.
authors.forename: Maria
authors.initials: M
authors.lastname: Ntoulia
authors.forename: Aliki
authors.initials: A
authors.lastname: Stathopoulou
authors.forename: Michail
authors.initials: M
authors.lastname: Ignatiadis
authors.forename: Nikos
authors.initials: N
authors.lastname: Malamos
authors.forename: Dimitris
authors.initials: D
authors.lastname: Mavroudis
authors.forename: Vassilis
authors.initials: V
authors.lastname: Georgoulias
authors.forename: Evi S
authors.initials: ES
authors.lastname: Lianidou
mesh_terms: D014408:Biomarkers, Tumor; D001943:Breast Neoplasms / Q000097:blood / Q000175:diagnosis*; D016022:Case-Control Studies; D045744:Cell Line, Tumor; D006801:Humans; D060785:Mammaglobin A; D009363:Neoplasm Proteins / Q000097:blood*; D009360:Neoplastic Cells, Circulating / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000097:blood*; D012334:RNA, Neoplasm / Q000097:blood*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014598:Uteroglobin / Q000097:blood*
chemical_list: D014408:Biomarkers, Tumor; D060785:Mammaglobin A; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D012334:RNA, Neoplasm; C556448:SCGB2A2 protein, human; D014598:Uteroglobin