Gene amplification and protein expression of EGFR and HER2 by chromogenic in situ hybridisation and immunohistochemistry in atypical adenomatous hyperplasia and adenocarcinoma of the lung.
Journal: 2005/November - Journal of Clinical Pathology
ISSN: 0021-9746
Abstract:
OBJECTIVE
To investigate the importance of gene amplification and EGFR (epidermal growth factor receptor) and HER2 protein expression during the progression of adenocarcinoma of the lung.
METHODS
EGFR and HER2 gene amplification was examined in atypical adenomatous hyperplasia (AAH), bronchioloalveolar carcinoma (BAC), and adenocarcinoma with mixed subtypes (MX) by chromogenic in situ hybridisation (CISH), and protein expression was examined by immunohistochemistry using paraffin wax embedded tissues.
RESULTS
EGFR and HER2 gene amplification was found in four and two of 86 cases, respectively, and was detected only in the invasive components of MX. EGFR and HER2 protein expression was seen in 24 and 18 of 86 cases, respectively. EGFR and HER2 proteins were not expressed in AAH but were expressed in one BAC case each. EGFR and HER2 proteins were expressed in 23 and 17 of 55 adenocarcinomas with MX. EGFR and HER2 protein expression was seen more often in the invasive components than in the BAC components of MX, and increased significantly as lesions progressed from AAH to BAC, early MX, and overt MX. Because EGFR and HER2 protein expression was frequently seen without gene amplification, other mechanisms apart from gene amplification may be associated with protein expression.
CONCLUSIONS
EGFR and HER2 gene amplification may be a late event and EGFR and HER2 protein expression may be associated with the development of adenocarcinoma of the lung.
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J Clin Pathol 58(10): 1076-1080

Gene amplification and protein expression of EGFR and HER2 by chromogenic in situ hybridisation and immunohistochemistry in atypical adenomatous hyperplasia and adenocarcinoma of the lung

Department of Pathology, Graduate School of Biomedical Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8551, Japan
Department of Molecular and Internal Medicine, Graduate School of Biomedical Sciences, Hiroshima University
Correspondence to:
Dr H Awaya
Department of Pathology, Graduate School of Biomedical Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8551, Japan; pj.ca.u-amihsorih@ianiok
Correspondence to:
Dr H Awaya
Department of Pathology, Graduate School of Biomedical Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8551, Japan; pj.ca.u-amihsorih@ianiok
Accepted 2005 Mar 3.

Abstract

Aims: To investigate the importance of gene amplification and EGFR (epidermal growth factor receptor) and HER2 protein expression during the progression of adenocarcinoma of the lung.

Methods: EGFR and HER2 gene amplification was examined in atypical adenomatous hyperplasia (AAH), bronchioloalveolar carcinoma (BAC), and adenocarcinoma with mixed subtypes (MX) by chromogenic in situ hybridisation (CISH), and protein expression was examined by immunohistochemistry using paraffin wax embedded tissues.

Results: EGFR and HER2 gene amplification was found in four and two of 86 cases, respectively, and was detected only in the invasive components of MX. EGFR and HER2 protein expression was seen in 24 and 18 of 86 cases, respectively. EGFR and HER2 proteins were not expressed in AAH but were expressed in one BAC case each. EGFR and HER2 proteins were expressed in 23 and 17 of 55 adenocarcinomas with MX. EGFR and HER2 protein expression was seen more often in the invasive components than in the BAC components of MX, and increased significantly as lesions progressed from AAH to BAC, early MX, and overt MX. Because EGFR and HER2 protein expression was frequently seen without gene amplification, other mechanisms apart from gene amplification may be associated with protein expression.

Conclusions: EGFR and HER2 gene amplification may be a late event and EGFR and HER2 protein expression may be associated with the development of adenocarcinoma of the lung.

Keywords: adenocarcinoma, chromogenic in situ hybridisation, epidermal growth factor receptor, HER2, lung
Abstract

Adenocarcinoma is the most frequent histological type of carcinoma of the lung. According to the World Health Organisation classification,1 atypical adenomatous hyperplasia (AAH) is a preneoplastic lesion of adenocarcinoma, and bronchioloalveolar carcinoma (BAC) is a non-invasive type of carcinoma. Adenocarcinoma with mixed subtypes (MX), which is the most common type of adenocarcinoma, usually includes BAC-type carcinoma in the periphery and invasive carcinoma in the central portion. Based on these morphological findings, it has been suggested that at least some adenocarcinomas occur as a result of progression from AAH through BAC to invasive adenocarcinoma.2–4

“Chromogenic in situ hybridisation allows comparison with histological findings obtained by microscopy”

Epidermal growth factor receptor EGFR/erbB1 and HER2/erbB2 are members of the EGFR family of type I receptor tyrosine kinases.5 Homodimerisation or heterodimerisation of these receptors appears to promote signal transduction pathways, including angiogenesis, inhibition of apoptosis, and cell growth. The EGFR gene is located on chromosome 7p12. The frequency of EGFR expression is high in non-small cell lung cancer (NSCLC). In previous studies, EGFR expression was detected in 38–75% of adenocarcinomas, 30–82% of squamous cell carcinomas, and 33% of large cell carcinomas.6–8 In contrast, the frequency of EGFR gene amplification, as determined by fluorescent in situ hybridisation (FISH), has been reported to be 9% in NSCLC.6 HER2, which is located on chromosome 17q21, is considered a heterodimerisation partner for other members of the HER family. Among NSCLC, HER2 expression is seen frequently in adenocarcinoma and large cell carcinoma and is associated with poor prognosis.9,10 In addition, HER2 expression was reported in 29–35% of adenocarcinomas, 1–18% of squamous cell carcinomas, and 0–20% of large cell carcinomas.10–13 The frequency of HER2 gene amplification, as determined by FISH, was reported to be 2–4% in NSCLC.11–13

Chromogenic in situ hybridisation (CISH) was introduced in 2000 and has been used to examine several types of carcinoma, such as breast carcinoma, prostate carcinoma, and extramammary Paget’s disease.14–18 However, there have been no previous reports of analysis of gene amplification in carcinoma of the lung using CISH. Although FISH is a standard technique for gene amplification, CISH has been reported to show a good correlation with gene amplification by FISH. Moreover, CISH allows comparison with histological findings obtained by microscopy.

In our present study, gene amplification and protein expression of EGFR and HER2 were examined by CISH in AAH, BAC, and MX, and the significance of the findings was analysed on the assumption of progression from AAH through BAC to invasive adenocarcinoma.

Take home messages

Abbreviations

  • AAH, atypical adenomatous hyperplasia

  • BAC, bronchioloalveolar carcinoma

  • CISH, chromogenic in situ hybridisation

  • EGFR, epidermal growth factor receptor

  • FISH, fluorescent in situ hybridisation

  • MX, adenocarcinoma with mixed subtypes

  • NSCLC, non-small cell lung cancer

Abbreviations

REFERENCES

REFERENCES

References

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