Evaluation of the epidermal growth factor receptor (EGFR) in colorectal tumours and lymph node metastases

doi:10.1016/S0959-8049(02)00234-4

European Journal of Cancer

Volume 38, Issue 17, November 2002, Pages 2258-2264

https://doi.org/10.1016/S0959-8049(02)00234-4 Get rights and content

Abstract

Overexpression of the epidermal growth factor receptor (EGFR) often correlates with an aggressive tumour phenotype and poor prognosis. To examine the relevance of EGFR in colorectal cancer, we determined the expression of EGFR protein in 249 colorectal adenocarcinomas and 42 lymph node metastases using immunohistochemistry. Moreover, we investigated a (CA)_n dinucleotide repeat polymorphism of the EGFR gene in a subset of 114 tumours. High levels of EGFR protein were observed in 123/249 (49.4%) samples. EGFR expression in colorectal carcinomas correlated with differentiation grade (P=0.014). However, there were no associations with Dukes' stage, site, patient age or gender. EGFR protein expression did not influence survival in this colorectal cancer patient cohort (P⩾0.05). Expression was not identical in paired colorectal tumours and lymph node metastases, with only 17/42 (40.5%) samples showing equivalent EGFR levels (P>0.05). The distribution of the (CA)_n dinucleotide repeat alleles in colorectal adenocarcinomas was not associated with EGFR protein expression (P>0.05). These results indicate that while EGFR overexpression is a common event in colorectal carcinogenesis, it does not influence patient prognosis.

Introduction

Colorectal cancer is the second most common malignancy in the developed world, which accounts for over 124 000 deaths per annum in Europe [1]. At present, patient prognosis is determined primarily by spread of the tumour at presentation. However, patients with an identical pathological disease stage can display widely differing outcomes in terms of survival and response to chemotherapy [2]. It is therefore fundamentally important to identify molecular markers of more aggressive colorectal tumour phenotypes in order to adjust patient treatment accordingly.

The epidermal growth factor receptor (EGFR, c-erbB-1) is the first identified member of the type I receptor tyrosine kinase family and is a major regulator of several distinct, diverse cellular pathways [3]. The ability to activate multiple pathways results from the numerous activating ligands which can bind EGFR and also from the various fellow family members with which EGFR can form dimer complexes [4]. Activated EGFR has been shown to cause transformation in vitro[5] and a wide range of human tumours have been shown to overexpress EGFR in vivo, including breast [6], lung [7], and bladder [8] carcinomas. While EGFR has been related to more aggressive behaviour in a number of tumour types [9], the clinical relevance of EGFR overexpression in colorectal cancer has not been comprehensively investigated.

Knowledge of EGFR levels within colorectal adenocarcinomas is also important in terms of therapeutic intervention. EGFR is the focus of various anticancer agents currently under investigation. Modulators of EGFR activity, for example anti-EGFR monoclonal antibodies and low molecular weight inhibitors of EGFR 10, 11, 12, have been shown to inhibit tumour proliferation in vitro and in certain cases promote apoptosis. Clinical trials are currently underway to assess the toxicities and anti-tumour activities of such compounds in vivo10, 11, 12.

While overexpression of EGFR can be attributed to gene amplification, this is more often not the case for solid tumours [13]. Although mutations in the EGFR gene are rarely observed, a polymorphic (CA)_n dinucleotide repeat in intron 1 of the EGFR gene has been reported [14] which is associated with gene expression [15]. In a model system, transcription was shown to have a stepwise decrease with increasing numbers of (CA)_n repeats (n=14 to n=21; [15]). In addition, further study has shown that the (CA)_n dinucleotide repeat length modulates intratumoral EGFR content in breast tumours in vivo[16]. However, to date there have been no studies examining the (CA)_n repeat in colorectal tumours.

This study has evaluated EGFR protein expression in a series of 249 primary colorectal adenocarcinomas and 42 corresponding lymph node metastases using immunohistochemistry, to investigate the influence of EGFR on colorectal cancer. In addition, we have ascertained the EGFR gene (CA)_n dinucleotide repeat polymorphism length in a subset of 114 primary colorectal tumours, to determine if it plays a role in this disease.

Section snippets

Patients and tumour samples

Archived tumour samples were available from 249 patients who had undergone elective surgery for colorectal cancer at Grampian University Hospitals NHS Trust, Scotland, UK, between 1994 and 1998. Lymph node metastases from a sub-group of 42 Dukes' C and D patients were also available. Samples were collected at the Department of Pathology, University of Aberdeen, under the auspices of the Aberdeen Colorectal Initiative. Tumours were routinely fixed in 10% neutral buffered formalin for 24 h, then

Immunohistochemistry—primary colorectal tumours

The expression of the EGFR protein was examined in 249 primary colorectal tumours, and the levels of expression are detailed in Table 1. One hundred and eighty-one tumours (72.7%) were found to express EGFR, with almost half of the tumours (123/249, 49.4%) displaying high levels of this protein (>50% of tumour cells positive). Of the 181 immunoreactive tumours, 73 displayed membranous EGFR expression, with a further 94 showing both membranous and cytoplasmic immunostaining. A small number of

Discussion

Overexpression of EGFR in vivo has been demonstrated in a number of human tumour types, including breast, lung, head/neck and bladder carcinomas 6, 7, 8, and has been related to a more aggressive clinical behaviour in a number of reports. While experimental data have demonstrated a direct relationship between the expression of EGFR by colorectal cancer cells and their ability to produce hepatic metastases [21], the role of EGFR overexpression in colorectal tumours in vivo has not been

Acknowledgments

This study was supported by a University of Aberdeen Development Trust Colorectal Cancer Initiative Grant (Steering Committee: Professor Jim Cassidy, Dr Howard L McLeod, Dr Graeme I. Murray, Professor Neva Haites, Professor Julian Little and Dr William T. Melvin), and a Grampian University Hospitals Trust Endowment Grant.

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Evaluation of the epidermal growth factor receptor (EGFR) in colorectal tumours and lymph node metastases

Abstract

Introduction

Section snippets

Patients and tumour samples

Immunohistochemistry—primary colorectal tumours

Discussion

Acknowledgments

Trends Pharmacol. Sci.

FEBS Lett.

Cell

Eur. J. Cancer

J. Biol. Chem.

Eur. J. Cancer

Estimates of the worldwide incidence of eighteen major cancers in 1985

Int J Cancer

Tumour markers of prognosis in colorectal cancer

Br. J. Cancer

Epidermal growth-factor receptor (EGFR)—results of a 6 year follow-up-study in operable breast cancer with emphasis on the node negative group

Br. J. Cancer

Immunohistochemical evidence of autocrine growth factors in adenocarcinoma of the lung

Cancer Res.

The epidermal growth factor receptor and the prognosis of bladder cancer

Cancer

Biochemical and clinical implications of the ErbB/HER signaling network of growth factor receptors

Adv. Cancer Res.