PMID- 15628907
OWN - NLM
STAT- MEDLINE
DCOM- 20050602
LR  - 20210527
IS  - 1543-2165 (Electronic)
IS  - 0003-9985 (Linking)
VI  - 129
IP  - 1
DP  - 2005 Jan
TI  - Copy number analysis of c-erb-B2 (HER-2/neu) and topoisomerase IIalpha genes in 
      breast carcinoma by quantitative real-time polymerase chain reaction using 
      hybridization probes and fluorescence in situ hybridization.
PG  - 39-46
AB  - CONTEXT: The Topoisomerase IIalpha (TOP2A) protein is the target of the 
      anthracycline class of chemotherapeutic agents. TOP2A is frequently coamplified 
      with c-erb-B2 and consequently might be a prognostic and/or predictive factor for 
      breast cancer patients when anthracycline-based chemotherapy is a consideration. 
      A total of 20% to 35% of breast carcinomas show amplification of the erb-B2 gene, 
      some of which also have coamplification of the TOP2A gene. Investigation of the 
      prognostic or predictive significance of these gene amplifications requires a 
      reliable and sensitive method for the measurement of gene copy number in clinical 
      tumor samples. OBJECTIVE: To assess 2 different assay methods that might allow 
      accurate, reproducible, quantitative, and high-throughput estimation of gene copy 
      number in fresh, frozen, or paraffin-embedded breast cancer specimens. DESIGN: We 
      developed an assay and analyzed the gene copy numbers of the erb-B2 and TOP2A 
      genes in 8 breast cancer cell lines, 6 fresh frozen samples, and 38 
      paraffin-embedded breast tumor specimens by a novel real-time polymerase chain 
      reaction (PCR) assay using hybridization probes. The results were compared with 
      standard fluorescence in situ hybridization. RESULTS: We discovered a 100% 
      concordance between assessment of gene copy number of erb-B2 and TOP2A between 
      quantitative PCR and fluorescence in situ hybridization (FISH). Quantitative PCR 
      also had the additional feature of uncovering an erb-B2 gene polymorphism. 
      Finally, we observed that TOP2A amplification only occurred in conjunction with 
      erb-B2 amplification in our paraffin-embedded cases of invasive breast carcinoma 
      and that this event was present in 5 (42%) of 12 erb-B2 amplified cases. 
      CONCLUSIONS: We conclude that the potentially automatic, real-time PCR analysis 
      using hybridization probes is an efficient method to perform copy number 
      analysis, with results that appear identical to the FISH technique and with the 
      benefit of identifying HER-2 polymorphisms.
FAU - Murthy, Sabita K
AU  - Murthy SK
AD  - Department of Pathology, The University of Calgary, Alberta, Canada.
FAU - Magliocco, Anthony M
AU  - Magliocco AM
FAU - Demetrick, Douglas J
AU  - Demetrick DJ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Arch Pathol Lab Med
JT  - Archives of pathology & laboratory medicine
JID - 7607091
RN  - 0 (Antigens, Neoplasm)
RN  - 0 (DNA Probes)
RN  - 0 (DNA, Neoplasm)
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Poly-ADP-Ribose Binding Proteins)
RN  - EC 5.99.1.3 (DNA Topoisomerases, Type II)
RN  - EC 5.99.1.3 (TOP2A protein, human)
SB  - IM
MH  - Antigens, Neoplasm/*genetics
MH  - Breast Neoplasms/*genetics/pathology
MH  - Carcinoma/*genetics/pathology
MH  - Cell Line, Tumor
MH  - Computer Systems
MH  - DNA Probes/genetics
MH  - DNA Topoisomerases, Type II/*genetics
MH  - DNA, Neoplasm/genetics
MH  - DNA-Binding Proteins/*genetics
MH  - Gene Amplification/*genetics
MH  - *Gene Dosage
MH  - Genes, erbB-2/*genetics
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Poly-ADP-Ribose Binding Proteins
MH  - Polymerase Chain Reaction/*methods
MH  - Predictive Value of Tests
MH  - Reproducibility of Results
MH  - Sensitivity and Specificity
EDAT- 2005/01/05 09:00
MHDA- 2005/06/03 09:00
CRDT- 2005/01/05 09:00
PHST- 2005/01/05 09:00 [pubmed]
PHST- 2005/06/03 09:00 [medline]
PHST- 2005/01/05 09:00 [entrez]
AID - OA4070 [pii]
AID - 10.5858/2005-129-39-CNAOCN [doi]
PST - ppublish
SO  - Arch Pathol Lab Med. 2005 Jan;129(1):39-46. doi: 10.5858/2005-129-39-CNAOCN.