title: Real-time RT-PCR: a complementary method to detect HER-2 status in breast carcinoma.
abstract: The option to treat patients presenting with HER-2 overexpressed invasive breast carcinoma with Herceptin requires quantitative determination of the HER-2 status. The aim of this study was to retrospectively evaluate the HER-2 mRNA expression levels using quantitative real-time RT-PCR (Q-RT-PCR) in tissue samples from 44 primary breast carcinomas and compare the results with immunohistochemistry (IHC). To determine the cut-off for altered mRNA expression, a normalized HER-2 expression value was determined for 20 normal breast tissue RNAs. Gene expression was categorized into three groups: normal expression (mRNAs <3); moderate overexpression (3 < or = mRNAs < 10) and strong overexpression (mRNAs < or =10). More than 38% (17/44) displayed strong overexpression, 25% (11/44) moderate and 36.3% (16/44) normal expression. Compared to IHC, only 7/44 cases were slightly discordant: strong mRNA overexpression/2+ protein staining (1 case), moderate overexpression/ 1+ (3 cases) and moderate overexpression/3+ (3 cases). These results show a high concordance rate (84%) between Q-RT-PCR and IHC (p < 10(-4)). We conclude that Q-RT-PCR is a useful complementary method for determination of the HER-2 status.
authors.affiliation: Département de Pathologie, Centre René Huguenin, Saint-Cloud, France. u539@nantes.inserm.fr
authors.forename: Céline
authors.initials: C
authors.lastname: Bossard
authors.forename: Ivan
authors.initials: I
authors.lastname: Bieche
authors.forename: Viviane
authors.initials: V
authors.lastname: Le Doussal
authors.forename: Rosette
authors.initials: R
authors.lastname: Lidereau
authors.forename: Jean Christophe
authors.initials: JC
authors.lastname: Sabourin
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism*; D005260:Female; D006801:Humans; D008875:Middle Aged; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000096:biosynthesis* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D019914:Taq Polymerase / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D018719:Erb-b2 Receptor Tyrosine Kinases; D019914:Taq Polymerase