title: Breast cancer-specific mRNA transcripts presence in peripheral blood after adjuvant chemotherapy predicts poor survival among high-risk breast cancer patients treated with high-dose chemotherapy with peripheral blood stem cell support.
abstract: OBJECTIVE
  To study the prognostic significance of the presence of breast cancer-specific mRNA transcripts in peripheral blood (PB), defined by serial analysis of gene expression, in high-risk breast cancer (HRBC) patients undergoing high-dose chemotherapy after receiving adjuvant chemotherapy.
  
  
  METHODS
  From 1994 to 2000, 84 HRBC patients (median age, 44 years; > 10 nodes; 74%) received adjuvant chemotherapy (fluorouracil, epirubicin, and cyclophosphamide for six cycles [83%] or doxorubicin and cyclophosphamide followed by paclitaxel) before undergoing one course of cyclophosphamide plus thiotepa plus carboplatin (STAMP V). Radiotherapy or hormone therapy was administered whenever indicated. Aliquots of apheresis-mononuclear blood cells were frozen from each patient. mRNA was isolated using an automatic nucleic acid extractor based on the magnetic beads technology; reverse transcription was performed using random hexamers. Cytokeratin 19, HER-2, P1B, PS2, and EGP2 transcripts were quantified to B-glucuronidase by real-time polymerase chain reaction (RT-PCR) using a linear DNA probe marked with a quencher and reporter fluorophores used in RT-PCR. Presence of PB micrometastases, estrogen receptor and progesterone receptor status, tumor size, age, tumor grade, number of nodes affected, and treatment with paclitaxel were included in the statistical analysis.
  
  
  RESULTS
  Median follow-up was 68.3 months (range, 6 months to 103 months). Forty-seven relapses (56%) and 35 deaths (41.7%) were registered. Both tumor size and presence of micrometastases reached statistical significance according to the Cox multivariate model. Relapse hazard ratio (HR) for those patients with PB micrometastases was 269% (P = .006); death HR, 300% (P = .011). Time relapse was 53 months longer for patients without micrometastases: 31.3 v 84.2 months (P = .021).
  
  
  CONCLUSIONS
  PB micrometastases presence after adjuvant chemotherapy predicts both relapse and death more powerful than classical factors in HRBC patients undergoing high-dose chemotherapy. Micrometastases search using a gene panel appears to be a more accurate procedure than classical approaches involving only one or two genes.
authors.affiliation: Medical Oncology Department, University Hospital 12 de Octubre, Madrid, Spain. quintelamiguel2000@yahoo.es
authors.forename: Miguel
authors.initials: M
authors.lastname: Quintela-Fandino
authors.forename: Joaquín Martínez
authors.initials: JM
authors.lastname: López
authors.forename: Ricardo
authors.initials: R
authors.lastname: Hitt
authors.forename: Soledad
authors.initials: S
authors.lastname: Gamarra
authors.forename: Antonio
authors.initials: A
authors.lastname: Jimeno
authors.forename: Rosa
authors.initials: R
authors.lastname: Ayala
authors.forename: Javier
authors.initials: J
authors.lastname: Hornedo
authors.forename: Cecilia
authors.initials: C
authors.lastname: Guzman
authors.forename: Florinda
authors.initials: F
authors.lastname: Gilsanz
authors.forename: Hernán
authors.initials: H
authors.lastname: Cortés-Funes
mesh_terms: D000328:Adult; D000368:Aged; D000954:Antigens, Surface / Q000235:genetics; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000008:administration & dosage / Q000627:therapeutic use*; D014408:Biomarkers, Tumor / Q000097:blood*; D001943:Breast Neoplasms / Q000097:blood / Q000737:chemistry / Q000235:genetics* / Q000473:pathology* / Q000628:therapy; D002135:Calcium-Binding Proteins / Q000235:genetics; D016190:Carboplatin / Q000008:administration & dosage; D017024:Chemotherapy, Adjuvant; D003520:Cyclophosphamide / Q000008:administration & dosage; D004317:Doxorubicin / Q000008:administration & dosage; D015251:Epirubicin / Q000008:administration & dosage; D000071858:Epithelial Cell Adhesion Molecule; D005260:Female; D005472:Fluorouracil / Q000008:administration & dosage; D005500:Follow-Up Studies; D006801:Humans; D007633:Keratins / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics; D008875:Middle Aged; D015999:Multivariate Analysis; D016017:Odds Ratio; D017239:Paclitaxel / Q000008:administration & dosage; D036102:Peripheral Blood Stem Cell Transplantation; D011237:Predictive Value of Tests; D053766:Presenilin-2; D011379:Prognosis; D012322:RNA Precursors / Q000097:blood*; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics; D012189:Retrospective Studies; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018570:Risk Assessment; D012307:Risk Factors; D016019:Survival Analysis; D013852:Thiotepa / Q000008:administration & dosage; D016896:Treatment Outcome; D000099211:Phospholamban
chemical_list: D000954:Antigens, Surface; D014408:Biomarkers, Tumor; D002135:Calcium-Binding Proteins; D016190:Carboplatin; D003520:Cyclophosphamide; D004317:Doxorubicin; D015251:Epirubicin; D000071858:Epithelial Cell Adhesion Molecule; D005472:Fluorouracil; D007633:Keratins; D008565:Membrane Proteins; D017239:Paclitaxel; D053766:Presenilin-2; D012322:RNA Precursors; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D018719:Erb-b2 Receptor Tyrosine Kinases; D013852:Thiotepa; D000099211:Phospholamban; C499379:EPCAM protein, human; C508861:PSEN2 protein, human