Epidermal growth factor receptor in ovarian tumours: correlation of immunohistochemistry with ligand binding assay

Abstract

Summary Epidermal growth factor receptor (EGFR) was studied in ovarian tumours with immunohistochemical (IH) and ligand-binding assay (LBA). Two different monoclonal antibodies (MoAbs: 2E9, EGFR1) with respect to detecting EGFR with different ligand-binding affinities (low, high and low) were used. When comparing the IH data of MoAbs 2E9 and EGFR1 a significant correlation was found (2P<0.0001). Both antibodies stained 77% of the adenocarcinoma samples. Evidence is increasing that growth factors and their receptors are involved not only in control of normal cell growth, but also in diseases, including cancer. The epidermal growth factor (EGF) and its receptor (EGFR) in particular have been investigated extensively in several tumour types. The EGFR molecule comprises a 170 kDa membrane protein, exhibiting an extracellular ligand (EGF or TGF-a) binding domain, a trans-membrane region and an intracellular domain facing the cytoplasm and exhibiting tyrosine kinase function. In many cell types the external domain displays high affinity (minor class) and low affinity (major class) EGF binding sites. The high-affinity binding sites prove to be most important for the activation of the signal transducing cascade (Defize et al., 1989). Both EGF and EGFR play an essential role in the development of mammary tissue (Tailor-Papadimitriou et al., 1977). Overexpression of EGFR in human primary breast cancer has been shown to be an indicator of a bad prognosis with respect to both relapse-free and overall survival (Sainsbury et al., 1987) and response to hormonal therapy of advanced disease (Nicholson et al., 1989). However, no consensus exists regarding the prognostic significance of EGFR . With respect to the ovary, changes in the level of EGF and EGFR in normal and neoplastic (benign/malignant) ovarian tissue specimens and its relation to clinical outcome have been studied less extensively (Bauknecht et al., , 1989(Bauknecht et al., , 1990Berchuck et al., 1991; Owens et al., 1991). In the present study we have investigated EGFR status in ovarian tissues with immunohistochemical (IH) and biochemical techniques (LBA: ligand binding assay). The IH technique was chosen for comparison with LBA because of its ability to identify tumour positivity at the cellular level, even in small tumour samples, excluding the influence of variance of tumour cellularity and the presence of EGFR in non-tumour tissue. Moreover, two different monoclonal antibodies (MoAbs: 2E9, EGFR1) were used to study possible differences in staining pattern between monoclonal antibodies reactive to different subtypes of receptor with respect to its ligand binding affinity. Materials and methods PatientsOne-hundred and twenty-eight tumours (121 epithelial and 7 non-epithelial), and 21 non-tumorous ovaries were analysed.Tumours were classifed in accordance with WHO classification (Serov et al., 1973). Forty-six patients (mean age 58 years) had a serous adenocarcinoma (37 primary, 9 metastatic), 20 patients (mean age 59 years) had a mucinous adenocarcinoma (20 prima...

Gonadotropins upregulate the epidermal growth factor receptor through activation of mitogen-activated protein kinases and phosphatidyl-inositol-3-kinase in human ovarian surface epithelial cells

“…The presence of EGFR in ovarian cancer has been well demonstrated using various methods such as ligand binding, immunohistochemistry, or Northern blot analysis (Bauknecht et al 1988, Battaglia et al 1989, Berchuck et al 1991, Morishige et al 1991, Owens et al 1991, HenzenLogmans et al 1992, Bauknecht et al 1993. EGFR is also frequently amplified and/or overexpressed when compared with normal OSE, and transfection with an antisense construct of EGFR into human ovarian cancer cell lines suppressed the malignant phenotype, cellular proliferation and tumorigenicity of these cells, suggesting its prognostic importance (Berns et al 1992, Brader et al 1998, Alper et al 2000. Furthermore, the contribution of a TGFa/EGF receptor autocrine loop to the growth of epithelial ovarian cancer cells is confirmed by several reports.…”

Expression of epidermal growth factor receptor in urinary bladder cancer metastases

“…Scale bar in (a), 500 µm; scale bar in (b), 25 µm. Macias, 1986;Battaglia, 1989;Toi, 1991;Henzen-Logmans, 1992) whereas others found no difference (Berchuck, 1992).…”

The relationship of quantitative epidermal growth factor receptor expression in non-small cell lung cancer to long term survival

“…It is, however, quantitative and we ensured to cut tumour tissue from the centre of the tumour to prepare membrane preparations. A significant correlation has been shown between maximum binding capacities of EGFr obtained from Scatchard plots and the percentage of positive tumour cells obtained by immunohistochemical staining with monoclonal antibody EGFR1 on ovarian carcinomas, Henzen-Logmans et al (1992).…”