Immunoreactive transforming growth factor alpha and epidermal growth factor in oral squamous cell carcinomas

Forty oral squamous cell carcinomas have been investigated immunohistochemically for the presence of transforming growth factor alpha (TGF-α) and epidermal growth factor (EGF). The same cases were recently characterized for the expression of EGF-receptors. TGF-α was detected with a monoclonal mouse antibody and EGF with polyclonal rabbit antiserum. Thirty-five of the tumours were positive for TGF-α and 26 of the tumours for EGF. None of the poorly differentiated tumours was positive for EGF, but they all were for TGF-α. In sections including normal differentiated oral mucosa, the cells above the basal cell layer were positive for both TGF-α and EGF. The same staining pattern was observed in oral mucosa obtained from healthy persons. In moderately to well differentiated carcinomas, the immunoreactivity was mainly confined to the cytologically more differentiated cells, thus paralleling the situation observed in the normal differentiated oral mucosa. In four cases, material was available from both a primary tumour and a metastasis. Three of these were positive for TGF-α and EGF with the same staining pattern as that of the primary tumours. This investigation together with our previous results confirms the existence of TGF-α, EGF, and EGF-receptors in the majority of oral squamous cell carcinomas and their metastases.     

Patterns of epidermal growth factor receptor amplification in malignant gliomas.

Amplification of the gene for epidermal growth factor receptor (EGFR) is a common finding in malignant gliomas. We found that 18 of 29 grade 3 and grade 4 gliomas had EGFR amplification when assayed using fluorescence in situ hybridization. The amplification pattern suggests that the amplicon is contained within double minute chromosomes in most cases. EGFR copy number can differ by 20-fold in amplified cells within a single case. Polysomy 7 occurs frequently in both EGFR-amplified and -unamplified cells. More than one-third of the cases had < or = 10 percent of cells with amplified EGFR, and it is likely that these cases would not have been identified by methods that do not examine DNA on a cell by cell basis.     

Immunohistochemical expression of transforming growth factor alpha and epidermal growth factor receptor in pancreatic endocrine tumors.

Coexpression of transforming growth factor alpha (TGF-alpha) and its receptor epidermal growth factor receptor (EGFR) is known to be associated with aggressive biologic behavior and adverse clinical outcome in a variety of tumors, including pancreatic adenocarcinomas. However, very little information is currently available as to whether this is true of pancreatic endocrine tumors (PETs) as well. Thirty-five PETs were retrospectively studied for immunohistochemical expression of TGF-alpha, the intracellular and extracellular domains of EGFR, and various hormonal secretory products. Proliferative activity was additionally studied (in 20 cases only) using the MIB-1 antibody. Thirty-one (89%) of 35 tumors were reactive for 1 or more of the peptide hormones tested; 22 (63%) tumors were positive for TGF-alpha; and 23 (65%) were positive for the intracellular and/or extracellular domain of EGFR. Based on their TGF-alpha and EGFR expression, these tumors could be classified into 4 groups. Of the 10 tumors in group I (positive for TGF-alpha and the complete EGFR molecule), 3 were malignant, 6 were >2 cm in diameter, 5 were functional, and 1 had a proliferative index of >40%. The 12 tumors in group II (positive for TGF-alpha but negative for the intracellular and/or extracellular domain of EGFR) included 4 malignant tumors, 4 PETs >2 cm in diameter, 8 functional, and 1 with a proliferative index of >40%. The 7 PETs in group III (positive for the intracellular/extracellular domain of EGFR alone) included 3 malignant tumors, 3 PETs >2 cm in diameter, and 3 functional tumors. The 6 tumors in group IV (completely negative for both TGF-alpha and EGFR) included 4 malignant tumors, 3 PETs >2 cm in diameter, and 4 functional lesions. Therefore, immunohistochemical expression of TGF-alpha and EGFR, either alone or in concert, shows no correlation with size, functional status, secretory profile, or biologic behavior and hence cannot be used as a marker of malignancy in this group of tumors.     

Immunohistochemical study of epidermal growth factor receptor expression in esophageal squamous cell carcinoma

Primary tumors from 31 patients with esophageal squamous cell carcinoma (ESCC) were immunohistochemically studied for the expression of markers of ESCC in order to define the clinical value of the levels of EGFR and HER-2 in the tumors. EGFR and HER-2 hyperexpression in the tumors of patients with ESCC was ascertained to be an important marker for the analysis of the clinical features of ESCC. There was an association of the elevated levels of EGFR and HER-2 in the tumors of ESCC patients with the presence of vascular tumor invasion (p = 0.006) and that with the poor outcome of the disease (p = 0.004). The findings suggest that estimation of changes found in EGRF and HER-2 expression in the tumors of patient with ESCC is of great interest for the individual prediction of the disease course and for the development of new approaches to treating these tumors, including targeted therapy against these tyrosine kinase receptors.     

Adhesion-mediated squamous cell carcinoma survival through ligand-independent activation of epidermal growth factor receptor

The survival and growth of squamous epithelial cells require signals generated by integrin-matrix interactions. After conversion to squamous cell carcinoma, the cells remain sensitive to detachment-induced anoikis, yet in tumor cell aggregates, which are matrix-deficient, these cells are capable of suprabasal survival and proliferation. Their survival is enhanced through a process we call synoikis, whereby junctional adhesions between neighboring cells generate specific downstream survival signals. Here we show that in squamous cell carcinoma cells, E-cadherin-mediated cell-cell contacts specifically induce activation of epidermal growth factor receptor (EGFR). EGFR activation in turn triggers the ERK/MAPK signaling module, leading to elevation of anti-apoptotic Bcl-2. After intercellular adhesion, formation of adherens junctions triggers the formation of E-cadherin-EGFR complexes, correlating with EGFR transactivation. Analysis of the process with a dominant-negative EGFR mutant indicated that activation of EGFR is ligand-independent. Our data implicate cell-cell adhesion-induced activation of EGFR as a cooperative mechanism that generates compensatory survival signaling, protecting malignant cells from detachment-induced death.     

Prognostic implications of epidermal growth factor receptor immunoreactivity in squamous cell carcinoma of the oral mucosa

To evaluate the clinical significance of the expression of epidermal growth factor receptor (EGFr) in oral squamous cell carcinoma (SCC), 100 formalin-fixed, paraffin-embedded cases of this tumour and ten samples of normal oral mucosa were immunostained with a monoclonal anti-EGFr antibody using an immunoalkaline phosphatase (APAAP) technique. EGFr immunoreactivity was detected in 36 of 100 tumours and in all samples of normal mucosa. Tumour cells demonstrated distinct membrane staining in 14 cases and predominantly cytoplasmic staining in 22 additional cases. EGFr was exclusively localized on the cell membrane of normal epithelial cells. Kaplan–Meyer survival curves and Cox proportional hazard regression models were used to assess overall survival and disease-free survival. A significant positive correlation was shown between EGFr membranous immunoreactivity and prolonged survival, in both univariate and multivariate analyses. Accordingly, patients with oral SCC showing down-regulated expression of membranous EGFr, who are more likely to suffer recurrence and death, should be strictly followed up and possibly treated with more aggressive therapeutic regimens. © 1998 John Wiley & Sons, Ltd.     

Immunohistochemical analysis of c-erbB-2 oncogene product and epidermal growth factor receptor expression in human urinary bladder carcinomas

Expression of the product of the c-erbB-2 gene, a proto-oncogene related to, but distinct from c-erbB-1 encoding the epidermal growth factor receptor (EGF-R), was investigated in human urinary bladder carcinomas. In addition, levels of EGF-R and transferrin receptor were also analyzed using an immunohistochemical approach, and the results compared with histological pattern and grading, and tumor staging. Increased expression of c-erb B-2 product was found in 32% of cases (7/22), a positive reaction being observed in 60% of transitional cell carcinoma (TCC) Grade 3 lesions (3/5), 20% of Grade 2 TCCs (2/10) and 100% of adenocarcinomas (AC) (2/2), but in none of the cases of squamous cell carcinoma (SCC). Although no statistical correlation with staging was evident, TCCs or SCCs of high grade and stage often showed EGF-R-positive staining, whereas other well differentiated lesions and normal bladder epithelium were generally negative. Most cases of urinary bladder carcinoma were positive for the transferrin receptor, which was not detected in normal bladder. The results thus suggested that a positive reaction for c-erbB-2 product is correlated with TCC histological grading or AC morphology. A high intensity of EGF-R staining in human bladder carcinomas may be associated with poor differentiation and invasion, whereas transferrin receptor expression might reflect tumor growth.     

Expression of epidermal growth factor receptor in squamous cell carcinomas of the anal canal is independent of gene amplification.

Immunohistochemical detection of expression of the epidermal growth factor receptor (EGFR) has been utilized to identify eligible patients with solid malignant tumors, including colorectal adenocarcinoma, for monoclonal antibody therapy (eg, cetuximab). The EGFR status in squamous cell carcinoma of the anal canal, an uncommon malignancy traditionally treated with chemoradiation, has not been well investigated. In this study, 38 primary squamous cell carcinomas of the anal canal were immunohistochemically examined for EGFR expression and analyzed by fluorescence in situ hybridization (FISH) for EGFR gene copy numbers. The results showed a variable degree of EGFR expression in 21 (55%) tumors, among which 13 (62%) cases exhibited a 2+ to 3+ staining pattern according to the Dako EGFR phamDx interpretation guide. There were no significant differences among tumors stratified by stage, degree of keratinization, or tissue block storage times. FISH analysis showed that none of the 34 cases with interpretable results had EGFR gene amplification. Increased gene copy numbers due to polysomy 7 were seen in seven of 18 (39%) cases that expressed EGFR protein and four of 16 (25%) cases that did not (P=0.3876). Ten (56%) tumors with positive EGFR staining showed a balanced disomy 7 pattern and one case with monosomy 7 exhibited strong EGFR expression (3+). These results demonstrate that EGFR is overexpressed in more than one-half of the squamous cell carcinomas of the anal canal through mechanisms other than gene amplification. These observations may have important therapeutic implications since EGFR-based targeted therapies have shown promise for other malignant neoplasms.     

Analysis of epidermal growth factor receptor and activated epidermal growth factor receptor expression in pituitary adenomas and carcinomas.

Epidermal growth factor receptor plays an important role in the pathogenesis of many malignancies. Various growth factors, including epidermal growth factor receptor, have been shown to influence pituitary tumor growth and differentiation. To analyze the role of epidermal growth factor receptor in pituitary tumor development, we examined normal pituitaries (n=8), pituitary adenomas (n=158), and pituitary carcinomas (n=7) for expression of epidermal growth factor receptor protein and messenger RNA using tissue microarrays and RT-PCR. We also examined (a) the expression of phospho-epidermal growth factor receptor, the activated form of epidermal growth factor receptor, in pituitary tumors and normal pituitaries by immunohistochemistry and (b) the effects on epidermal growth factor receptor expression of treating pituitary cells (HP75 cell line) with epidermal growth factor. Epidermal growth factor receptor and the phosphorylated variant expression were present in normal pituitary cells. Epidermal growth factor receptor messenger RNA was also detected in normal pituitaries, pituitary adenomas, and carcinomas by in situ hybridization and RT-PCR. Most pituitary adenomas showed expression of epidermal growth factor receptor and the phosphorylated variant. Nonfunctional adenomas showed higher levels of expression of epidermal growth factor receptor (76 vs 34%) and of phospho-epidermal growth factor receptor (26 vs 8%) as compared to functional adenomas. Five of seven pituitary carcinomas showed strong expression of both epidermal growth factor receptor and phospho-epidermal growth factor receptor. When a human pituitary cell line (HP75) was cultured in the presence of epidermal growth factor receptor, there was an increase in the levels of both epidermal growth factor receptor and phospho-epidermal growth factor receptor after 5 h of treatment, thus confirming that epidermal growth factor receptor signaling was active in pituitary tumors. These results indicate that activated epidermal growth factor receptor is expressed in pituitary adenomas and carcinomas. Higher levels in pituitary carcinomas suggest a role in pituitary tumor progression.     

Glandular squamous cell carcinoma of the urinary bladder

The paper gives a clinical observation of a 52 year-old male with a rare histological urinary bladder tumor primary grandular-squamous-cell carcinoma (pT3N IM0). The tumor is represented by two components large acinic-cell adenocarcinoma and squamous-cell carcinoma with keratinization, which smoothly pass one into another; the tumor has grown through all layers of the urinary bladder wall but it has failed to grow into the peritoneum. A microscopic study has indicated that the urachus is intact. Metastases were found in 3 of 8 lymph nodes: one showed high-grade adenocarcinoma and two others displayed average-grade squamous-cell carcinoma.     

Immunocytochemical study of epidermal growth factor receptor, transforming growth factor alpha, and “squamous differentiation” in human endometrial carcinoma

The expression of epidermal growth factor receptor (EGFr) and transforming growth factor alpha (TGF-α) was compared with the presence of “squamous differentiation” (SD) visualized in various histotypes of endometrial carcinoma by using a panel of monoclonal antibodies. The results of the current study demonstrate that EGFr and TGF-α are present in routinely processed endometrial carcinoma. The highest positive EGFr and TGF-α expression was seen in the group of adenocarcinomas with SD. The more intense EGFr and TGF-α immunoreactivity was observed in “squamous” foci both in adenoacanthomas (AA) and in adenosquamous carcinomas (AS). These EGFr- and TGF-α-positive squamous areas prevalently displayed a “stratification-related” cytokeratin (CK) immunoprofile characterized by the expression of CKs 1, 4, 5, 10, 13, 14, and 16. No correlation was found between EGFr- and TGF-α-positive status and depth of myometrial invasion or surgical stage. These results clearly demonstrate that EGFr and TGF-α expression is related remarkably to endometrial carcinoma with “squamous” areas both morphologically and immunophenotypically. This specific association leads us to suggest that EGFr and TGF-α expression in endometrial carcinoma may be prevalently involved in the equilibrium of cell differentiation of the “squamous” foci commonly observed in this group of neoplasias.     

Expression of neu protein, epidermal growth factor receptor, and transforming growth factor alpha in breast cancer. Correlation with clinicopathologic parameters.

The major objectives of this study were twofold: to determine 1) if growth factors or growth factor receptors were expressed similarly or differently in a clinically well-characterized group of breast cancer patients and 2) if these phenotypic characteristics were associated with any of the commonly used prognostic parameters. Formalin-fixed paraffin-embedded tumor tissue from 51 node-positive breast cancer patients were analyzed for the expression of neu, epidermal growth factor-receptor (EGF-R), and transforming growth factor alpha (TGF alpha) using immunoperoxidase staining. Positive membranous staining for neu was observed in 15 (29%) tumors. Over-expression of neu was observed in high-grade, estrogen-receptor-negative tumors (P less than 0.05). Epidermal growth factor receptor was expressed in 22 (43%) of the tumors analyzed and found to a greater degree in estrogen-receptor-negative and high-grade tumors (P less than 0.025). A significant correlation between neu and EGF-R expression was also noted. Tumors expressing membranous staining of neu had a greater than 70% chance of expressing EGF-R (P less than 0.01). Expression of TGF alpha was found in 68% of tumors and TGF alpha was detected in grade 1 and 2 tumor to a greater degree than EGF-R. The authors conclude that assaying tumors for these antigens may give additional phenotypic characteristics that can give further insight into the biology of breast cancer.     

Basaloid squamous cell carcinoma occurring in the urinary bladder

Basaloid squamous cell carcinoma is a recently described, distinct variant of squamous cell carcinoma that arises predominantly in the upper aerodigestive tract. Herein we report a case of basaloid squamous cell carcinoma arising in the urinary bladder. The patient was a 60-year-old woman who experienced intractable urinary tract infections following multiple corrective surgical procedures for incontinence. Biopsies of cystoscopically evident flat lesions were performed, and the patient subsequently underwent a radical cystectomy. Histologically, the lesions consisted of nests of basaloid cells with brisk mitotic activity, areas of squamous differentiation along with areas of squamous metaplasia, and squamous cell carcinoma in situ. These features are similar to those of basaloid squamous cell carcinoma described elsewhere in the body. To our knowledge, this is the first reported case of basaloid squamous cell carcinoma in the urinary bladder.     

Immunohistochemical localisation of epidermal growth factor, transforming growth factor alpha and EGF receptor during organogenesis of the murine hypophysis in vivo

In previous study of melanocyte-stimulating hormone (MSH) cell development in the proliferating pars intermedia, which is in close apposition to the presumptive pars nervosa, no direct cell-to-cell contact was found between the boundary neurohypophyseal pituicytes (PIC), adenohypophyseal precursor stem cells (PSC) and the related diencephalic mesenchymal cells. Here, we have used immunohistochemistry to examine cytokine expression in the development of the hypophysis during foetal stages II-IV. Light and confocal laser scanning microscopy indicated diffuse expression of both TGF alpha and EGF in the hypophysis at different foetal stages. While no findings indicative for temporary changes of TGF alpha and EGF patterns were found in the foetal hypophysis, a temporary increment of EGF molecules was distinct in the diencephalic mesenchyme at stages III and IV. On the other hand, light microscopy intensively immuno-localized EGFR in the adenohypophysis and neurohypophysis at different developmental stages. Immunoreactivity of EGFR in the cytoplasm and nucleus suggested active proliferative events in the PIC and PSC of stages II-IV mouse pituitaries.     

Spindle cell lesions of the bladder and urinary tract

Spindle cell lesions of the urinary tract encompass a variety of benign and malignant tumours as well as a group of lesions of controversial nomenclature that is the subject of ongoing debate. This review focuses on our current and evolving understanding of the lesion variably referred to as inflammatory myofibroblastic tumour, pseudosarcomatous myofibroblastic proliferation or inflammatory pseudotumour and the main differential diagnoses of sarcomatoid carcinoma and sarcoma. Other spindle cell lesions of the bladder are described.     

Immunodetection of the ligand-activated receptor for epidermal growth factor.

Many receptors for cellular growth factors are known to be protein tyrosine kinases which become activated upon ligand binding at their extracellular domain. We describe here a method to detect the activation state of Epidermal Growth Factor receptor (EGFr) with a monoclonal antibody (mAb74). This antibody was found to preferentially recognize the ligand-activated EGFr as detected by immunoprecipitation, Western blotting and immunocytochemical techniques. mAb74 did not recognize other tyrosine-phosphorylated proteins and was not inhibited by phosphotyrosine, suggesting that it is recognizing an epitope specific for the ligand-activated EGF receptor. The reactivity of mAb74 towards EGFr was closely correlated with the EGF-dependent tyrosine phosphorylation of endogenous substrates. This antibody allows one to detect the activated EGF receptor in vitro or in vivo even in a complex mixture of other tyrosine kinases and substrates.     

A comparison of epidermal growth factor receptor (EGFR) and c-erbB-2 oncogene expression in head and neck squamous cell carcinomas.

The proto-oncogenes c-erbB-2 and epidermal growth factor (EGF) receptor which encode 2 closely homologous transmembrane glycoproteins have been found amplified and/or overexpressed in a range of epithelial malignancies. In a series of 46 head and neck squamous cell cancers (SCCs), immunohistochemical reactivity for the EGF receptor was detected in all cases, particularly at the invading edge of cellular islands of SCC and in the basal cells of normal adjacent squamous epithelium. Southern blot analysis demonstrated EGF receptor gene amplification in 3 cases. In contrast, strong membrane staining for the c-erbB-2 oncoprotein was not detected in any sample, and there were no cases of c-erbB-2 gene amplification. Despite a close structural and (presumed) functional homology between these 2 receptor-oncoproteins in the development of malignancy, we report that their expression in SCCs is markedly different. Furthermore, unlike the situation for breast cancer, quantitation of the c-erbB-2 or EGF receptor oncoproteins is unlikely to yield important prognostic information in this group of patients.