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Updated: Aug 11, 2026
Heterogeneity Mapping of Protein Expression in Tumors using Quantitative Immunofluorescence
Published on: October 25, 2011
Pitfalls in immunohistochemical assessment of EGFR expression in soft tissue sarcomas
C Kersting1, J Packeisen, B Leidinger
1Institute of Pathology, University of Münster, Münster, Germany.
Background:
New targeted cancer treatments acting against growth factor receptors such as the epidermal growth factor receptor (EGFR) necessitate selecting patients for treatment with these drugs. Besides carcinomas, soft tissue sarcomas (STS) express EGFR and might thereby be a promising target for this new therapeutic strategy.
Objective:
To test and compare different EGFR antibodies to determine the frequency of EGFR expression in STS.
Methods:
302 consecutive specimens of STS were examined using the tissue microarray technique. EGFR expression levels were assessed by immunohistochemistry using five different commercially available antibodies. Gene amplification status was measured by fluorescence in situ hybridisation (FISH). Immunoreactivity and amplification status were correlated with clinicopathological features and follow up data available in 163 cases.
Results:
EGFR expression frequency ranged between 0.3% and 52.9%, depending on the antibody and scoring method used. In all, 3.5% of the tumours showed egfr gene amplification by FISH, which correlated with EGFR expression for three antibodies. Only one antibody had independent prognostic value in multivariate analysis and correlated with an unfavourable outcome; egfr gene amplification status showed no correlation with clinical features.
Conclusions:
Frequency of EGFR immunopositivity in STS strongly depends on the antibody used, and only one of five antibodies tested predicted an unfavourable clinical outcome. This indicates that choice of primary antibody and scoring system have a substantial impact on the determination of EGFR immunoreactivity.
Insights
The choice of antibody significantly impacts epidermal growth factor receptor (EGFR) detection in soft tissue sarcomas (STS). Only one of five tested antibodies predicted unfavorable outcomes, highlighting the need for careful antibody selection in EGFR-targeted therapy for STS.
Area of Science:
- Oncology
- Pathology
- Molecular Biology
Background:
- Targeted cancer therapies, including those against the epidermal growth factor receptor (EGFR), require precise patient selection.
- Soft tissue sarcomas (STS) express EGFR, making them a potential target for novel therapeutic strategies.
Purpose of the Study:
- To evaluate and compare the efficacy of different EGFR antibodies in determining EGFR expression frequency within STS.
- To assess the correlation between EGFR expression, gene amplification, and clinicopathological features in STS.
Main Methods:
- Utilized tissue microarray technique on 302 STS specimens.
- Assessed EGFR expression via immunohistochemistry with five distinct commercial antibodies.
- Measured gene amplification status using fluorescence in situ hybridisation (FISH).
Main Results:
- EGFR expression varied widely (0.3%–52.9%) based on the antibody and scoring method.
- EGFR gene amplification was detected in 3.5% of tumors, correlating with EGFR expression for three antibodies.
- One antibody demonstrated independent prognostic value, indicating an unfavorable outcome, while gene amplification showed no clinical correlation.
Conclusions:
- The frequency of EGFR immunopositivity in STS is highly dependent on the chosen antibody.
- Only one of the five tested antibodies accurately predicted an unfavorable clinical outcome.
- Antibody selection and scoring systems significantly influence the determination of EGFR immunoreactivity in STS.
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