title: Quantitative Analysis of ErbB1 and ErbB2 Genes Amplification by a High Performance Liquid Chromatography.
abstract: BACKGROUND
  Genes for human epidermal growth factor receptors B1 (ErbB1) and B2 (ErbB2) were amplified in breast and ovarian cancers. Both of them were associated with aggressive disease and worse prognosis. The ErbB1 or ErbB2 status of a tumor may provide an indication of the response to ErbB1 and ErbB2 -targeted therapies. For accurate and rapid assessment of amplification of ErbB1 and ErbB2 oncogenes, a High Performance Liquid Chromatography (HPLC) method was developed in this study.
  
  
  METHODS
  DNA was extracted from 30 primary breast tumors and 20 blood samples of healthy donors. ErbB1 and ErbB2 genes along with a reference gene were co-amplificated by Polymerase Chain Reaction (PCR). The PCR products were separated and quantified using an anion-exchange column within 30 min and in a single step. Optimum resolution was obtained when a sodium chloride gradient and a column temperature of 35°C were used. The results of HPLC analysis of ErbB1 and ErbB2 PCR products were compared with real time PCR method as a gold standard test for 7 tumor samples.
  
  
  RESULTS
  The proposed HPLC method was confirmed by real time PCR method. Twenty two and ten of the specimens in our breast cancer cohort showed more than a two-fold amplification of ErbB2 and ErbB1 oncogenes, respectively.
  
  
  CONCLUSIONS
  Our results were confirmed by real time PCR and showed that HPLC method is a specific, cheap and clinically applicable analytical approach for assessment of ErbB1 and ErbB2 statuses in breast tumors.
authors.affiliation: Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
authors.forename: Mozhgan
authors.initials: M
authors.lastname: Rasti
authors.affiliation: Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
authors.forename: Zohreh
authors.initials: Z
authors.lastname: Honardar
authors.affiliation: Department of Biochemistry, School of Medicine, Yasuj University of Medical Sciences, Yasuj, Iran.
authors.forename: Mohsen
authors.initials: M
authors.lastname: Nikseresht
authors.affiliation: Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
authors.forename: Aliakbar
authors.initials: A
authors.lastname: Owji
keywords: Epidermal growth factor; Gene amplification; High pressure liquid chromatography; Methods; Polymerase chain reaction