Skip to main content

Quality control of immunohistochemical assay of HER-2/neu expression

Context

HER-2/neu (c-erb-2) gene encodes a membrane receptor related to the epidermal growth factor receptor and its amplification has been correlated with a shorter disease free interval in breast cancer patients. Recently, clinical trials have shown that treatment with an antibody can block growth of cells expressing HER-2/neu and prolong survival of patients. An immunohistochemical (IHC) test and fluorescence in situ hybridisation (FISH) are used to predict HER-2/neu status. IHC is economic to use and included in routine diagnostic services. However, IHC sensitivity varies between laboratories. In order to ensure reproducibility of results a standard control is needed so that day-to-day variation can be monitored. The objective of this study was to develop a control for HER-2/neu histochemical detection by investigating four breast and ovarian cell lines.

Significant findings

A 100% agreement between testing centres using FISH revealed that MDA-MB-453 and SKOV-3 cell lines showed HER-2/neu amplification, while BT-20 and MCF-7 were negative. SKOV-3 exhibited a score of 3+ HER-2/neu protein overexpression by IHC and gene amplification with FISH; in contrast, MCF-7 cells showed neither protein expression nor gene amplification (0 score). For BT-20 cells an 86% concordance (6/7) was observed between a 0 or 1+ score by IHC and no amplification detected by FISH, while 71% concordance between an HIC score of 2+ and amplification detected by FISH was shown for MDA-MB-453. The sensitivity of the developed cell-line control was 3+, 2+, 2+ and 0 with CB11 and 3+, 2+, 1+, 0 with the HercepTest or DAKO antibody (SKOV-3, MDA-MB-453, BT-20 and MCF-7 respectively).

Comments

Composite blocks were produced, providing controls for immunohistochemical analysis of HER-2/neu. These blocks were tested alongside commercially available kits in seven cancer centres in the UK and France. The developed control has the advantage of providing a graded series of expression for each of the 3+, 2+, 1+ and 0 categories and can be implemented in the routine clinical setting. As a first step this control will help monitoring HER-2/neu assay sensitivity between various laboratories, despite the use of an antibody and antigen retrieval system. However, the development of a cell-line microarray in the near future will be advantageous in assisting with HER2/neu diagnosis.

Methods

Cell culture, IHC, FISH, antibodies (CB11 clone, HercepTest and DAKO polyclonal)

Additional information

Schnitt SJ and Jacobs TW: Current status of HER2 testing: caught between a rock and a hard place. Am J Clin Pathol 2001, 116: 806-810. (PubMed)

References

  1. 1.

    Rhodes A, Jasani B, Couturier J, McKinley M, Morgan J, Dodson A, Navabi H, Miller K, Balaton A: A formalin-fixed paraffin-processed cell line standard for quality control of immunohistochemical assay of HER-2/neu expression in breast cancer. Am J Clin Pathol. 2002, 117: 81-89.

    CAS  Article  PubMed  Google Scholar 

Download references

Author information

Affiliations

Authors

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Skliris, G. Quality control of immunohistochemical assay of HER-2/neu expression. Breast Cancer Res 4, 76400 (2002). https://doi.org/10.1186/bcr-2002-76400

Download citation

Keywords

  • Breast and ovarian cell lines, FISH, HER-2/neu, immunohistochemistry