PMID- 19309522
OWN - NLM
STAT- MEDLINE
DCOM- 20090624
LR  - 20211020
IS  - 1471-2407 (Electronic)
IS  - 1471-2407 (Linking)
VI  - 9
DP  - 2009 Mar 23
TI  - Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise 
      methods to detect HER-2 status in breast carcinoma.
PG  - 90
LID - 10.1186/1471-2407-9-90 [doi]
AB  - BACKGROUND: HER-2 gene testing has become an integral part of breast cancer 
      patient diagnosis. The most commonly used assay in the clinical setting for 
      evaluating HER-2 status is immunohistochemistry (IHC) and fluorescence in situ 
      hybridization (FISH). These procedures permit correlation between HER-2 
      expression and morphological features. However, FISH signals are labile and fade 
      over time, making post-revision of the tumor difficult. CISH (chromogenic in situ 
      hybridization) is an alternative procedure, with certain advantages, although 
      still limited as a diagnostic tool in breast carcinomas. METHODS: To elucidate 
      the molecular profile of HER-2 status, mRNA and protein expression in 75 invasive 
      breast carcinomas were analyzed by real time quantitative RT-PCR (qRT-PCR) and 
      IHC, respectively. Amplifications were evaluated in 43 of these cases by CISH and 
      in 11 by FISH. RESULTS: The concordance rate between IHC and qRT-PCR results was 
      78.9%, and 94.6% for qRT-PCR and CISH. Intratumoral heterogeneity of HER-2 status 
      was identified in three cases by CISH. The results of the three procedures were 
      compared and showed a concordance rate of 83.8%; higher discordances were 
      observed in 0 or 1+ immunostaining cases, which showed high-level amplification 
      (15.4%) and HER-2 transcript overexpression (20%). Moreover, 2+ immunostaining 
      cases presented nonamplified status (50%) by CISH and HER-2 downexpression 
      (38.5%) by qRT-PCR. In general, concordance occurred between qRT-PCR and CISH 
      results. A high concordance was observed between CISH/qRT-PCR and FISH. 
      Comparisons with clinicopathological data revealed a significant association 
      between HER-2 downexpression and the involvement of less than four lymph nodes (P 
      = 0.0350). CONCLUSION: Based on these findings, qRT-PCR was more precise and 
      reproducible than IHC. Furthermore, CISH was revealed as an alternative and 
      useful procedure for investigating amplifications involving the HER-2 gene.
FAU - Rosa, Fabiola E
AU  - Rosa FE
AD  - Department of Genetics, Institute of Biosciences, UNESP, Sao Paulo State 
      University, Botucatu, Sao Paulo, Brazil. fabiolarosa7@yahoo.com.br
FAU - Silveira, Sara M
AU  - Silveira SM
FAU - Silveira, Cassia G T
AU  - Silveira CG
FAU - Bergamo, Nadia A
AU  - Bergamo NA
FAU - Neto, Francisco A Moraes
AU  - Neto FA
FAU - Domingues, Maria A C
AU  - Domingues MA
FAU - Soares, Fernando A
AU  - Soares FA
FAU - Caldeira, Jose R F
AU  - Caldeira JR
FAU - Rogatto, Silvia R
AU  - Rogatto SR
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090323
PL  - England
TA  - BMC Cancer
JT  - BMC cancer
JID - 100967800
RN  - EC 2.7.10.1 (Receptor, ErbB-2)
SB  - IM
MH  - Adult
MH  - Aged
MH  - Aged, 80 and over
MH  - Breast Neoplasms/genetics/metabolism/*pathology
MH  - Female
MH  - Gene Amplification
MH  - Gene Dosage
MH  - Humans
MH  - Immunohistochemistry
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Middle Aged
MH  - Neoplasm Staging
MH  - Receptor, ErbB-2/*genetics/metabolism
MH  - Reproducibility of Results
MH  - Reverse Transcriptase Polymerase Chain Reaction/*methods
PMC - PMC2667535
EDAT- 2009/03/25 09:00
MHDA- 2009/06/25 09:00
PMCR- 2009/03/23
CRDT- 2009/03/25 09:00
PHST- 2008/06/11 00:00 [received]
PHST- 2009/03/23 00:00 [accepted]
PHST- 2009/03/25 09:00 [entrez]
PHST- 2009/03/25 09:00 [pubmed]
PHST- 2009/06/25 09:00 [medline]
PHST- 2009/03/23 00:00 [pmc-release]
AID - 1471-2407-9-90 [pii]
AID - 10.1186/1471-2407-9-90 [doi]
PST - epublish
SO  - BMC Cancer. 2009 Mar 23;9:90. doi: 10.1186/1471-2407-9-90.