PMID- 15494857
OWN - NLM
STAT- MEDLINE
DCOM- 20050322
LR  - 20170214
IS  - 1066-8969 (Print)
IS  - 1066-8969 (Linking)
VI  - 12
IP  - 4
DP  - 2004 Oct
TI  - Real-time PCR quantification of c-erbB-2 gene is an alternative for FISH in the 
      clinical management of breast carcinoma patients.
PG  - 311-8
AB  - Evaluation of gene amplification and protein expression of the c-erbB-2/neu in 
      breast carcinomas has been an important task in breast cancer management. 
      Although immunohistochemistry is widely applied, fluorescence in situ 
      hybridization (FISH) technology shows its advantage in discriminating tumors in 
      an objective manner. More recently, development of LightCycler technology permits 
      evaluation of gene amplification with a small volume of DNA run in a 20 microL 
      glass capillary. In this study, a series of 87 breast carcinomas were chosen for 
      evaluation of c-erbB-2/neu gene amplification detected by both LightCycler 
      technology and FISH. Real-time polymerase chain reaction (PCR) was performed in 
      LightCycler capillaries with 10 ng sample DNA. By using LightCycler Relative 
      Quantification Software version 1 (LightCycler, Roche, Mannheim, Germany), the 
      amount of c-erbB-2 DNA was calculated as a ratio of c-erbB-2/reference gene 
      quantity in a sample, and then the ratio was divided by the ratio of c-erbB-2 
      gene/reference gene quantities of a calibrator DNA (a standard DNA provided in 
      the kit), which was run with each sample reaction in parallel. Dual-color FISH 
      was performed on sections of the formalin-fixed, paraffin-embedded tissue array 
      samples using the DAKO HER2 FISH pharmDX kit (DAKO A/S, Glostrup, Danmark) 
      according to the manufacturer's instructions. Furthermore, immunohistochemistry 
      was performed in parallel, with both the NCL-CB11 and HercepTest antibodies. Both 
      the FISH technology and the LightCycler-PCR identified a similar percentage of 
      tumors with c-erbB-2 gene amplification in our present study, 16% (14/87) and 15% 
      (13/87), respectively, whereas immunohistochemistry demonstrated 32% and 34% 
      c-erbB-2 overexpression with the NCL-CB11 and HercepTest antibodies, 
      respectively. In addition, FISH and PCR were highly correlated in detecting 
      tumors mainly with 3+++ c-erbB-2 protein expression by immunohistochemistry, 
      indicating that LightCycler real-time quantification of c-erbB-2 gene may be an 
      alternative to FISH in breast cancer clinical application.
FAU - Suo, Zhenhe
AU  - Suo Z
AD  - Department of Pathology, The Norwegian Radium Hospital, University of Oslo, 
      Kristiansand, Norway.
FAU - Daehli, Kathinka U
AU  - Daehli KU
FAU - Lindboe, Christian Fr
AU  - Lindboe CF
FAU - Borgen, Elin
AU  - Borgen E
FAU - Bassarova, Assia
AU  - Bassarova A
FAU - Nesland, Jahn M
AU  - Nesland JM
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Int J Surg Pathol
JT  - International journal of surgical pathology
JID - 9314927
RN  - 0 (Biomarkers, Tumor)
RN  - 0 (DNA, Neoplasm)
RN  - 0 (Oncogene Proteins v-erbB)
SB  - IM
MH  - Biomarkers, Tumor/metabolism
MH  - *Breast Neoplasms/genetics/metabolism/pathology/therapy
MH  - Carcinoma/genetics/metabolism/pathology/therapy
MH  - DNA, Neoplasm/analysis
MH  - Female
MH  - Fluorescent Antibody Technique, Direct
MH  - *Genes, erbB-2
MH  - Humans
MH  - *In Situ Hybridization, Fluorescence
MH  - Light
MH  - Oncogene Proteins v-erbB/genetics/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction/*methods
MH  - Tissue Array Analysis
EDAT- 2004/10/21 09:00
MHDA- 2005/03/23 09:00
CRDT- 2004/10/21 09:00
PHST- 2004/10/21 09:00 [pubmed]
PHST- 2005/03/23 09:00 [medline]
PHST- 2004/10/21 09:00 [entrez]
AID - 10.1177/106689690401200404 [doi]
PST - ppublish
SO  - Int J Surg Pathol. 2004 Oct;12(4):311-8. doi: 10.1177/106689690401200404.