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Annals of Oncology Advance Access originally published online on March 9, 2007
Annals of Oncology 2007 18(5):845-850; doi:10.1093/annonc/mdm059
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© 2007 European Society for Medical Oncology

breast cancer

Quantitative real-time PCR analysis and microarray-based RNA expression of HER2 in relation to outcome

J Bergqvist1,*, JF Ohd1, J Smeds1, S Klaar1, J Isola2, H Nordgren3, GP Elmberger1, H Hellborg4, J Bjohle1, A-L Borg1, L Skoog1 and J Bergh1

1 Department of Oncology and Pathology, Karolinska Institute and University Hospital, Stockholm, Sweden
2 Laboratory of Cancer Genetics, Institute of Medical Technology and University Hospital of Tampere, Tampere, Finland
3 Department of Pathology, Uppsala University Hospital, Uppsala
4 Stockholm-Gotland Regional Tumor Registry, Oncology Centre, Karolinska University Hospital, Stockholm, Sweden

* Correspondence to: Dr J. Bergqvist, Department of Oncology and Pathology, Radiumhemmet, Karolinska Institute and University Hospital, SE-171 76 Stockholm, Sweden. Tel: +46-8-51776279; Fax: +46-8-51779524; E-mail: jenny.bergqvist{at}ki.se

Background: Our aim was to use quantitative real-time PCR (Q-PCR) and RNA expression profiles (RNA-EPs) to investigate HER2 status in relation to outcome.

Patients and methods: Cut-off levels for Q-PCR and RNA-EP were established in relation to immunohistochemistry (IHC) validated by FISH in a test set of frozen tissue samples from 40 primary breast cancers. The HER2 status was subsequently studied in another validation set of 306 tumors, where Q-PCR and RNA-EP results were compared with previously carried out IHC that we had validated by chromogenic in situ hybridization (CISH).

Results: Q-PCR and RNA-EP offered similar sensitivity (90% versus 77%), specificity (93% versus 95%), and negative (99% versus 98%) and positive (63% versus 61%) predictive values for HER2 determinations. Analyses of relapse-free survival (RFS) and overall survival on the basis of 5 and 10 years of follow-up indicated equivalent hazard ratios for all three techniques. In contrast to IHC/CISH, both Q-PCR and RNA-EP analyses of HER2 also gave statistically significant results regarding RFS and breast cancer-corrected survival after 10 years of follow-up.

Conclusion: The use of RNA-EP and Q-PCR to analyze HER2 in frozen and formalin-fixed breast cancer samples may be an alternate approach to IHC in combination with FISH/CISH.

Key words: breast cancer, CISH, diagnostic methods, HER2, quantitative real-time PCR, RNA expression profiles


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