title: Clinical evaluation of developed PCR-based method with hydrolysis probes for TOP2A copy number evaluation in breast cancer samples.
abstract: OBJECTIVE
  The objective of this study was to develop a new real time PCR-based method for quantitative detection of topoisomerase II alpha (TOP2A) aberrations and to evaluate its clinical utility in breast cancer.
  
  
  METHODS
  The method applied dually labelled hydrolysis probes and Pfaffl quantification method. The study group consisted of 83 consecutive breast cancer patients.
  
  
  RESULTS
  In the examined tumour samples median TOP2A gene dosage was 1.08 (range 0.34-7.55). TOP2A amplifications were found in 12 tumours (14.5%), no deletion was detected. Statistically significant positive correlation of TOP2A gene dosage with nodal status, tumour grade, and HER2 protein status was found. TOP2A status also correlated with disease free survival.
  
  
  CONCLUSIONS
  The newly developed real time PCR assay showed to be fast and easy to perform. Determined by the method TOP2A gene dosage was shown to be a potent prognostic factor in breast cancer.
authors.affiliation: Department of Medical Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdańsk and Medical University of Gdańsk, Gdańsk, Poland. azaczek@gumed.edu.pl
authors.forename: Anna
authors.initials: A
authors.lastname: Zaczek
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Markiewicz
authors.forename: Janusz
authors.initials: J
authors.lastname: Jaśkiewicz
authors.forename: Tadeusz
authors.initials: T
authors.lastname: Pieńkowski
authors.forename: Piotr
authors.initials: P
authors.lastname: Rhone
authors.forename: Jacek
authors.initials: J
authors.lastname: Jassem
authors.forename: Marzena
authors.initials: M
authors.lastname: Wełnicka-Jaśkiewicz
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000951:Antigens, Neoplasm / Q000235:genetics*; D001943:Breast Neoplasms / Q000235:genetics*; D004250:DNA Topoisomerases, Type II / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics*; D005260:Female; D018628:Gene Dosage / Q000235:genetics*; D006801:Humans; D006868:Hydrolysis; D008875:Middle Aged; D015345:Oligonucleotide Probes / Q000235:genetics*; D000075223:Poly-ADP-Ribose Binding Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
chemical_list: D000951:Antigens, Neoplasm; D004268:DNA-Binding Proteins; D015345:Oligonucleotide Probes; D000075223:Poly-ADP-Ribose Binding Proteins; D004250:DNA Topoisomerases, Type II; C000618811:TOP2A protein, human