title: Detection of circulating tumor cells in breast cancer patients using multiplex reverse transcription-polymerase chain reaction and specific primers for MGB, PTHRP and KRT19 correlation with clinicopathological features.
abstract: OBJECTIVE
  The aim of this study was to correlate the clinicopathological features of breast cancer patients with the positive detection of parathyroid hormone-related protein (PTHRP), cytokeratin protein 19 (KRT19) and mammaglobin (MGB) using a multiplex reverse transcription polymerase chain reaction (RT-PCR) assay developed to detect circulating tumor cells (CTCs) in peripheral blood of patients with breast cancer.
  
  
  METHODS
  Peripheral blood samples were collected from 54 breast cancer patients and 20 healthy blood donors. Subsequently, the samples were processed for RNA extraction and analyzed for the expression of PTHRP, KRT19 and MGB using specific primers and multiplex RT-PCR.
  
  
  RESULTS
  The positive detection rates in breast cancer patients for PTHRP, KRT19 and MGB were 68.5%, 63% and 22.2% and for healthy donors 10%, 0% and 10%, respectively. The statistical analysis revealed that PTHRP- and KRT19-positive detections correlated with the diagnosis of breast cancer while the combined positive detections of PTHRP-plus-KRT19 correlated with the presence of distant metastasis, especially with bone metastasis. Moreover, positive detections of KRT19 correlated with high proliferation rate of breast cancer tumors. MGB-positive detections did not add any diagnostic advantage in such analysis.
  
  
  CONCLUSIONS
  Multiplex-PCR based detection of CTCs using PTHRP and KRT19 primers can provide useful information for the disease.
authors.affiliation: Department of Experimental Physiology, National and Kapodistrian University of Athens, Medical School, Athens, Greece Second Department of Internal Medicine University of Athens, Hippokratio Hospital, Athens, Greece rikaskondra@gmail.com.
authors.forename: Maria
authors.initials: M
authors.lastname: Skondra
authors.affiliation: Department of Experimental Physiology, National and Kapodistrian University of Athens, Medical School, Athens, Greece.
authors.forename: Eliona
authors.initials: E
authors.lastname: Gkioka
authors.affiliation: Second Department of Propedeutic Surgery National and Kapodistrian University of Athens, Medical School, Athens, Greece.
authors.forename: Ioannis D
authors.initials: ID
authors.lastname: Kostakis
authors.affiliation: Department of Experimental Physiology, National and Kapodistrian University of Athens, Medical School, Athens, Greece.
authors.forename: Nikolaos
authors.initials: N
authors.lastname: Pissimissis
authors.affiliation: Department of Experimental Physiology, National and Kapodistrian University of Athens, Medical School, Athens, Greece.
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Lembessis
authors.affiliation: Second Department of Internal Medicine University of Athens, Hippokratio Hospital, Athens, Greece.
authors.forename: Dimitrios
authors.initials: D
authors.lastname: Pectasides
authors.affiliation: Department of Experimental Physiology, National and Kapodistrian University of Athens, Medical School, Athens, Greece.
authors.forename: Michael
authors.initials: M
authors.lastname: Koutsilieris
mesh_terms: D000328:Adult; D000368:Aged; D014408:Biomarkers, Tumor; D001859:Bone Neoplasms / Q000097:blood / Q000235:genetics / Q000556:secondary; D001943:Breast Neoplasms / Q000097:blood / Q000235:genetics / Q000473:pathology*; D018270:Carcinoma, Ductal, Breast / Q000097:blood / Q000235:genetics / Q000556:secondary*; D018275:Carcinoma, Lobular / Q000097:blood / Q000235:genetics / Q000556:secondary*; D016022:Case-Control Studies; D005260:Female; D005500:Follow-Up Studies; D006801:Humans; D053539:Keratin-19 / Q000235:genetics*; D008207:Lymphatic Metastasis; D060785:Mammaglobin A / Q000235:genetics*; D008875:Middle Aged; D060787:Neoplasm Grading; D009367:Neoplasm Staging; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D011379:Prognosis; D012333:RNA, Messenger / Q000097:blood / Q000235:genetics; D012334:RNA, Neoplasm / Q000097:blood / Q000235:genetics; D044168:Receptor, Parathyroid Hormone, Type 1 / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D014408:Biomarkers, Tumor; D053539:Keratin-19; D060785:Mammaglobin A; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D044168:Receptor, Parathyroid Hormone, Type 1; C556448:SCGB2A2 protein, human
keywords: KRT19; MGB; PCR; PTHRP; breast cancer; circulating tumor cells (CTCs)