title: Assessment of HER-2 gene overexpression in Isfahan province breast cancer patients using Real Time RT-PCR and immunohistochemistry.
abstract: OBJECTIVE
  Overexpression of proto-oncogene HER-2 is one of the main molecular markers of breast cancer involved in prognosis and diagnosis and also in trastuzumab therapy. Thus, a request for the evaluation of HER-2 status in breast cancer has been increasing. The aim of our study was assessment of HER-2 overexpression in malignant and benign breast cancer specimens by Real Time RT-PCR technique and comparison of its results with IHC outcomes.
  
  
  METHODS
  Twenty benign and sixty malignant breast cancers in addition to fifteen normal breast tissue specimens were analyzed by Real Time RT-PCR method. Fresh tissue samples were disrupted by mortar and pestle. A syringe and a needle were used for complete homogenization of the tissues. The RNA was then isolated from the samples and converted to cDNA. A standard curve was initially plotted using BioEasy SYBR Green I and then all 95 specimens were studied by Real Time RT-PCR using 2(-ΔΔCt) method.
  
  
  RESULTS
  23.3% of 60 malignant specimens showed HER-2 overexpression, while all of the benign samples represented the normal expression level of HER-2 gene. The concordance rate between the results of Real Time RT-PCR and IHC was 86.6%.
  
  
  CONCLUSIONS
  Real Time RT-PCR method is an almost reliable technique and at least can be used as a complementary method for confirming IHC results. This is emanated from relatively high rate of concordance between outcomes of IHC test, as a routine method of detecting the HER-2 gene expression status, and Real Time RT-PCR technique.
authors.affiliation: Genetics Division, Biology Department, Faculty of Sciences, University of Isfahan, Isfahan, Iran. Electronic address: Hosein.tabatabaeian@gmail.com.
authors.forename: Hosein
authors.initials: H
authors.lastname: Tabatabaeian
authors.forename: Zohreh
authors.initials: Z
authors.lastname: Hojati
mesh_terms: D000328:Adult; D000368:Aged; D001943:Breast Neoplasms / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D007492:Iran; D008875:Middle Aged; D000090063:Proto-Oncogene Mas; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics* / Q000378:metabolism*
chemical_list: D000090063:Proto-Oncogene Mas; D012333:RNA, Messenger; D018719:Erb-b2 Receptor Tyrosine Kinases; C000715470:MAS1 protein, human
keywords: Breast cancer; EGFR; ER; Epidermal growth factor receptor; Estrogen Receptor; FISH; Fluorescence in-situ hybridization; GAPDH; Gene overexpression; Glyceraldehyde-3-phosphate dehydrogenase; HER-2; IHC; ISH; Immunohistochemistry; In-situ hybridization; PR; Progesterone Receptor; Real Time RT-PCR; Real Time Reverse Transcriptase-PCR; Real Time Reverse Transcriptase-Polymerase Chain Reaction; SD; Standard deviation