epidermal-growth-factor and Salivary-Gland-Neoplasms

Expression of erbB2 and/or epidermal growth factor receptor (EGFR) is associated with biologic aggressiveness and poor prognosis in malignant salivary gland tumors (MSGTs). This phase II study was conducted to determine the antitumor activity of lapatinib, a dual inhibitor of EGFR and erbB2 tyrosine kinase activity, in MSGTs.. Patients with progressive, recurrent, or metastatic adenoid cystic carcinoma (ACC) immunohistochemically expressing at least 1+ EGFR and/or 2+ erbB2 were treated with lapatinib 1,500 mg daily, in a two-stage cohort. Patients with non-ACC MSGTs were treated as a separate single-stage cohort.. Of 62 patients screened, 29 of 33 (88%) ACC and 28 of 29 (97%) non-ACC patients expressed EGFR and/or erbB2. Forty patients with progressive disease were enrolled onto the study. Among 19 assessable ACC patients, there were no objective responses, 15 patients (79%) had stable disease (SD), nine patients (47%) had SD > or = 6 months, and four patients (21%) had progressive disease (PD). For 17 assessable non-ACC patients, there were no objective responses, eight patients (47%) had SD, four patients (24%) had SD > or = 6 months, and nine patients (53%) had PD. The most frequent adverse events were grade 1 to 2 diarrhea, fatigue, and rash. Eight paired tumor biopsies for correlative studies were procured; results did not correlate with clinical outcome.. Although no responses were observed, lapatinib was well tolerated, with prolonged tumor stabilization of > or = 6 months in 36% (95% CI, 21% to 54%) of assessable patients. The antitumor effects of lapatinib in MGSTs appear mainly cytostatic, hence evaluation of other molecular targeted agents, or combinations with lapatinib, may be considered. Continued efforts should be made to gain better understanding into the biology of this heterogeneous group of malignancies.

Topics: Adaptor Proteins, Signal Transducing; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Adenoid Cystic; Disease-Free Survival; Epidermal Growth Factor; Female; Follow-Up Studies; Humans; Lapatinib; Male; Middle Aged; Neoplasm Recurrence, Local; Protein-Tyrosine Kinases; Quinazolines; Salivary Gland Neoplasms; Treatment Outcome

Salivary adenoid cystic carcinoma (SACC) is one of the most common malignant cancers of the salivary gland, and 32.4-72.0% of SACC cases exhibit neural invasion (NI); however, the molecular mechanism underlying the high invasion potential of SACC remains unclear.. The present study investigated the role of epidermal growth factor receptor (EGFR) in the AKT inhibition- or mitogen-activated protein kinase kinase (MEK)-induced NI and epithelialmesenchymal transition (EMT) in SACC cells using EGFR, PI3K, and MEK inhibitors. SACC-83 cell viability was assessed using an MTT assay, and a wound healing assay was performed to evaluate cell migration. Immunohistochemical staining with streptavidin peroxidase was used to detect the positive expression rate of EMT, AKT, phosphorylated (p)-AKT, ERK, and p-ERK proteins. The impact of EGFR, PI3K, and MEK inhibitors on tumor growth and NI was examined in a xenograft model in nude mice.. EGF and EGFR are effective in increasing cell viability, migration, and invasion. SACC metastasis is affected by the PI3K/AKT and MEK/ERK pathways, both of which are initiated by EGF/EGFR. The EMT and NI are regulated by the EGF/EGFR, PI3K/AKT, and MEK/ERK pathways. The present findings demonstrate the importance of suppressed EGFR/AKT/MEK signaling in NI in SACC by neural-tumor co-culture in vitro. Furthermore, our preclinical experiment provides solid evidence that injection of EGFR, PI3K, and MEK inhibitors suppressed the tumor growth and NI of SACC cells in nude mice.. It was identified that inhibitors of EGFR, PI3K/AKT or MEK/ERK suppressed the proliferation, migration, and NI of SACC-83 cells via downregulation of the PI3K/AKT or MEK/ERK pathways. It was also demonstrated that inhibition of EGFR abolishes EMT in SACC by inhibiting the signaling of PI3K/AKT and MEK/ERK. The present results suggest the potential effectiveness of targeting multiple oncogenes associated with downstream pathways of EGF/EGFR, as well as potential therapeutic targets to limit NI in SACC by PI3K/AKT or MEK/ERK inhibition.

Topics: Animals; Carcinoma, Adenoid Cystic; Cell Line, Tumor; Cell Movement; Cell Proliferation; Epidermal Growth Factor; ErbB Receptors; Humans; Mice; Mice, Nude; Mitogen-Activated Protein Kinase Kinases; Neoplasm Invasiveness; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Salivary Gland Neoplasms

The aim of the present study was to investigate the expression of claudin-1, -3, -4, -5 and -7 proteins in mucoepidermoid carcinoma of oral cavity and analyze whether EGF may interfere in the expression of the genes that encode claudins using in vitro models.. Using immunohistochemistry, the expression of claudins was searched in 36 histologically graded cases of mucoepidermoid carcinoma. The association of expression of claudins with clinical-pathological parameters was evaluated. An in vitro step investigated the influence of EGF on gene expression of claudins by real time RT-PCR technique.. Claudin-1, -3, -4, -5, and -7 were highly expressed in most mucoepidermoid carcinomas. These expressions were compared with clinicopathological parameters. High expression of claudin-1 was associated with patients over 40 years-old (p = 0.05) and Caucasians (p = 0.024). In vitro experiments demonstrated a tendency for Claudin gene expression increase after EGF stimulus.. The expression of claudins is maintained in mucoepidermoid carcinoma cells and EGF could be related with this expression. Our results point out to a fundamental biological importance to CLDNs in normal and neoplastic tissue. The expression patterns of CLDNs does not yet allow a clinical application, but the biological knowledge will ground evidence to new studies towards possible target-therapies.

Topics: Adult; Biomarkers, Tumor; Carcinoma, Mucoepidermoid; Claudins; Epidermal Growth Factor; Female; Gene Expression Regulation, Neoplastic; Humans; Male; Middle Aged; Salivary Gland Neoplasms

During tumor development, benign neoplastic cells are influenced by the expression of cytokines, growth factors, and proteases present in the tumor microenvironment. Epidermal growth factor (EGF) is the most studied growth factor and is considered important for cell proliferation and migration. Metalloproteinases (MMPs) are also involved in tumor progression. The present study aimed to analyze the proliferation, viability and migration index of pleomorphic adenoma myoepithelial cells, in addition to the secretion of MMPs with EGF supplementation. Benign myoepithelial cells were cultured with two different EGF doses (5 and 10 ng/ml), and the influence of EGF on cell proliferation and viability, using trypan blue and MTT assays, respectively, after 24, 48, and 72 h, was evaluated. To analyze cellular morphology, hematoxylin-eosin staining and indirect immunofluorescence using the anti-vimentin antibody, was performed. In vitro migration assays were performed in Transwell chambers with an 8-μm pore covered with Matrigel and supplemented with 5 or 10 ng/ml of EGF, after 96 h. After 4 days of cell culture, ELISA was performed to determine the MMP-2 and MMP-13 levels. One-way analysis of variance (ANOVA) with post hoc Tukey test was applied, with a significance level of 0.05. The results revealed that EGF influences myoepithelial cell morphology, without alteration of proliferation and viability. The migration assay showed that EGF increased the mean index from 16 % in the control group to 40 and 76 % for 5 and 10 ng/ml of EGF, respectively. ELISA revealed that when the cells were supplemented with either of the EGF doses, an increase in MMP-2 levels was observed when compared with the control group (C). This study concludes that EGF aids in the production of MMP-2, which favors the dissolution of the basement membrane, contributing to cell migration and tumor progression, hence permitting contact between the myoepithelial cells and stroma.

Topics: Adenoma, Pleomorphic; Cell Movement; Cell Proliferation; Cell Shape; Epidermal Growth Factor; Epithelial Cells; Humans; Matrix Metalloproteinase 13; Matrix Metalloproteinase 2; Myoepithelioma; Salivary Gland Neoplasms; Signal Transduction; Tumor Cells, Cultured

Adenoid cystic carcinoma (ACC) is a relatively rare epithelial tumour of the salivary glands in the maxillofacial region. About 40-60% of the patients develop distant metastases, which have been documented most commonly in the lung but also in brain, bone, liver, thyroid, spleen and pancreatic gland.. A 55-year-old women with intraosseous ACC in the mandible mimicking apical periodontitis following curative resection and radiotherapy is presented. Three years later, multiple lung metastases were observed followed by chemotherapy. Five years after curative resection, the patient presented simultaneously with new expansive soft tissue in the pancreas and mammary gland as well as in the kidney found to be metastatic ACC. No case has been reported to date on the manifestation of distant metastases of intraosseous ACC in the breast and the kidney as described by these observations. Metastatic mammary gland ACC stained positive for epithelial growth factor receptor (EGFR) but was negative for HER-2/neu and Cyclooxygenase-2 (COX-2) expression.

Topics: Alveolar Bone Loss; Breast Neoplasms; Carcinoma, Adenoid Cystic; Cyclooxygenase 2; Diagnosis, Differential; Diagnostic Errors; Epidermal Growth Factor; Female; Humans; Hyperbaric Oxygenation; Kidney Neoplasms; Mandibular Neoplasms; Middle Aged; Osteomyelitis; Pancreatic Neoplasms; Periapical Periodontitis; Receptor, ErbB-2; Salivary Gland Neoplasms

Hematogenous metastasis is one of the most important factors determining the outcome of the patients with salivary adenoid cystic carcinoma (SACC). In the present study, we examined expression profile of genes in SACC cell lines to look for molecules responsible for its unique metastatic trait. A transcriptomic microarray analysis between the lower lung-metastatic rate cell line SACC-83 and the higher lung-metastatic rate cell line SACC-LM were performed, and eight genes, showed by microarray to be highly expressed in SACC-LM, were picked for validation by quantitative real-time PCR. Among the genes, the expression of epiregulin, a novel member of epidermal growth factor family, was 350-folds higher in SACC-LM than in SACC-83. Accordingly, we examined the effects of epiregulin on migration and invasion in SACC-83 as well as its targeted downstream molecules, and found that epiregulin could promote in vitro migration and invasion in SACC-83. Furthermore, epiregulin not only induced activation of both ERK1/2 and Akt, but also expression of COX-2. In addition, all these effects could be partially blocked by U0126, a specific inhibitor of mitogen-activated protein kinase kinase (MEK or MAPKK), or LY294002, a specific inhibitor of phosphatidylinositol 3-kinase (PI3K). Conclusively, the results suggest that epiregulin may play an important role in lung metastasis of SACC.

Topics: Carcinoma, Adenoid Cystic; Cell Line, Tumor; Cell Movement; Cyclooxygenase 2 Inhibitors; Epidermal Growth Factor; Epiregulin; Humans; Lung Neoplasms; Microarray Analysis; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Neoplasm Invasiveness; Proto-Oncogene Proteins c-akt; Reverse Transcriptase Polymerase Chain Reaction; Salivary Gland Neoplasms

To analyze the expression of ErbB-1 (Her-1 or EGFR), ErbB-2 (Her-2 or neu), ErbB-3 (Her-3) and ErbB-4 (Her-4) and their correlation in 3 different types of salivary gland tumors.. Immunohistochemical expression of epidermal growth factor (EGF), EGFR, ErbB-2, fatty acid synthase (FAS) and Ki-67 were analyzed in 41 pleomorphic adenoma (PA), in 30 mucoepidermoid carcinoma (MEC) and in 30 adenoid cystic carcinoma (ACC) and correlated with their histologic patterns.. EGF was more common in MEC and PA, but MEC had a higher percentage of strongly positive cases. EGFRc and EGFRm were both more frequent in MEC and ACC. Higher scores of ErbB-2c were observed in PA, followed by MEC and ACC. In contrast, higher scores of ErbB-2m were more common in MEC as compared to ACC and PA. FAS was most commonly found in PA and MEC. Moreover, MEC showed the highest percentage of strongly positive cases. Ki-67 was higher in MEC and ACC than in PA. From a correlation of immunomarkers with the histologic patterns, it was observed that cribriform ACC presented more expression of EGFR and high grade MEC showed a higher percentage of ErbB-2, FAS and Ki-67.. EGF, EGFR, ErbB-2 and FAS were commonly found and seem to be important in the tumorigenesis of salivary gland tumors, particularly in percentage of strongly positive cases. Ki-67 was higher in MEC and ACC than in PA. From a correlation of immunomarkers with the histologic patterns, it was observed that cribriform ACC presented more expression of EGFR and high grade MEC showed a higher percentage of ErbB-2, FAS and Ki-67.. EGF, EGFR, ErbB-2 and FAS were commonly found and seem to be important in the tumorigenesis of salivary gland tumors, particularly in MEC. (Anal Quant Cytol Histol 2009;31:280-287)

Topics: Adenoma, Pleomorphic; Biomarkers, Tumor; Carcinoma, Adenoid Cystic; Carcinoma, Mucoepidermoid; Epidermal Growth Factor; ErbB Receptors; fas Receptor; Female; Fluorescent Antibody Technique, Indirect; Humans; Immunoenzyme Techniques; Ki-67 Antigen; Male; Middle Aged; Receptor, ErbB-2; Salivary Gland Neoplasms

Topics: Biomarkers, Tumor; Carcinoma; Epidermal Growth Factor; ErbB Receptors; Humans; Immunoenzyme Techniques; Salivary Gland Neoplasms; Salivary Glands

This study used an immunohistochemical technique to assess the expression of epidermal growth factor (EGF) in 40 specimens of salivary adenoid cystic carcinoma (ACC), 7 specimens of labial glands adjacent to mucocele, and 5 specimens of normal submandibular glands. In normal submandibular glands, immunohistochemically detectable EGF was demonstrated in all ductal segments, including intercalated, striated, and excretory duct cells. No EGF positive staining was found in acinar compartments. including serous and mucous acinar cells. In degenerated labial glands adjacent to mucocele, no EGF staining was detected in the remaining acinar and ductal cells. In salivary ACCs, positive EGF immunostaining was observed in one of the 5 (20%) ACCs with a solid pattern and in 13 of the 35 (37.1%) ACCs with a tubular-cribriform pattern. The overall EGF expression rate in 40 salivary ACCs was 35%. Positive EGF staining was predominantly found in tubular structures in the tubular ACCs and in duct-like structures in large cribriform patterns or in the stroma of the cribriform ACCs. There was no significant correlation between EGF expression in salivary ACCs and any of the clinicopathological parameters including patient age and sex, cancer location, TNM status, clinical stage, histologic type, perivascular or perineural invasion, focal necrosis of tumor, and cellular atypia. We conclude that the duct segments of the normal submandibular gland are the sites of EGF synthesis and secretion. In degenerated labial glands adjacent to mucocele, EGF synthesis is completely inhibited. Furthermore, EGF is mainly biosynthesized in cells forming tubular or duct-like structures in tubular or cribriform salivary ACCs, and EGF may play a biologic role, particularly as a mitogen in salivary ACC growth.

Topics: Adult; Aged; Carcinoma, Adenoid Cystic; Epidermal Growth Factor; Female; Humans; Immunohistochemistry; Male; Middle Aged; Salivary Gland Neoplasms

We examined the proliferative signal transduction by EGF in HSG-AZA 3, a subclone of HSG cell line. The treatment of cells with EGF resulted in an increase in [3H]thymidine incorporation into DNA depending upon EGF concentrations. In addition, the nuclear proto-oncogene c-fos was rapidly induced by EGF. Moreover, EGF induced transient expression of EGF receptor mRNA followed by the de novo synthesis of EGF receptor protein. On the other hand, treatment of the cells with EGF occurred phosphorylation by tyrosine kinase comprised in the EGF receptor, autophosphorylation, followed by activation of MAP kinase. These results indicate that the proliferative response to EGF is modulated by the phosphorylation cascade mediating EGF receptor-associated tyrosine kinase and MAP kinase, and transient activation of c-fos protein is implicated in the cell proliferation.

Topics: Adenocarcinoma; Base Sequence; Calcium-Calmodulin-Dependent Protein Kinases; Cell Division; Epidermal Growth Factor; ErbB Receptors; Humans; Molecular Sequence Data; Phosphorylation; Proto-Oncogene Mas; Proto-Oncogene Proteins c-fos; Receptor Protein-Tyrosine Kinases; RNA, Messenger; Salivary Gland Neoplasms; Signal Transduction; Tumor Cells, Cultured

The adenocarcinoma cell line derived from an intercalated ductal epithelium of a human salivary gland (HSG) proliferates autonomously mediated by an epidermal growth factor-(EGF)-like molecule with a molecular weight of 46 kDa and an EGF receptor (EGFR). The c-erbB2 protein, a member of EGFR family was also expressed in HSG cells and was involved in the growth signal pathway of HSG cells as well as EGFR. The autocrine growth is regulated by glucocorticoid and retinoic acid (RA) via their receptors. Retinoic acid receptor (RAR) of HSG cells revealed a transcriptional activity in vivo, and the heterodimerization between RAR and 9-cis retinoic acid receptor (RXR) is requisite for the binding with a specific DNA element termed RA response element in vitro. RXR alpha and RXR beta were cloned from HSG cells, and these RXRs, together with RAR, seemed to play a physiological role in RA signaling in vivo.

Topics: Adenocarcinoma; Bucladesine; Cell Division; Cell Transformation, Neoplastic; Epidermal Growth Factor; Humans; Receptors, Retinoic Acid; Salivary Gland Neoplasms; Signal Transduction; Transcription, Genetic; Tretinoin; Tumor Cells, Cultured

We have previously shown that conditioned medium (CM) from metastasizing human salivary gland adenocarcinoma cell clones contains factor(s) that stimulate the proliferation and migration of bovine aortic endothelial (BAE) cells, and inhibit the production of collagenases by BAE cells (Azuma M. et al. (1993) Cancer Lett., 73, 85-93). To further characterize this, we evaluated the expression level of epidermal growth factor (EGF) secreted by a non-metastasizing cell clone (HSGc) and its metastasizing cell clones, and analysed the effect of EGF on the biologic behaviors of BAE cells. When the secretion of EGF by cell clones was estimated by enzyme-linked immunosorbent assay, metastasizing cell clones released a large amount of EGF as compared with HSGc. However, the number of EGF receptor was detected consistently at a level that was similar in all cell clones. With regard to the effect of EGF on the malignant potential of cell clones such as proteolytic aggressiveness, EGF did not affect the secretion of both collagenases and their inhibitor from cell clones. Alternatively, exogenous EGF stimulated the proliferation and migration of BAE cells, and inhibited the secretion of collagenases from BAE cells. Neutralization with a neutralizing antibody of EGF released into CM abolished the inhibitory effect of CM on the secretion of collagenases from BAE cells. Thus, the CM-contained factor, which is responsible for the induction of biologic behaviors of BAE cells, can be attributed to EGF.

Topics: Adenocarcinoma; Animals; Aorta; Cattle; Cell Division; Collagenases; Culture Media, Conditioned; Endothelium, Vascular; Epidermal Growth Factor; Glycoproteins; Humans; Matrix Metalloproteinase Inhibitors; Neoplasm Metastasis; Salivary Gland Neoplasms; Tissue Inhibitor of Metalloproteinases; Tumor Cells, Cultured

Human epidermal growth factor (hEGF) stimulates the growth and differentiation of various tissues. We measured EGF levels in saliva (n = 128), urine (n = 94), and serum (n = 99) with radioimmunoassay in order to study the kinetics of hEGF in saliva of normal subjects and patients with oral disease. Salivary EGF levels showed an apparent diurnal rhythm related to the taking of meals. Urinary and serum EGF levels showed no obvious diurnal rhythm. There was no significant correlation between salivary and urinary EGF levels, nor between salivary and serum EGF levels. Salivary EGF levels were significantly lower in the younger group (0-9 years old, 3.06 +/- 0.32 ng/ml, p < 0.05) than in the elder group (10-79 years old, 4.78 +/- 3.5 ng/ml), but did not correlate with age in the elder group. There was no significant difference between males and females between EGF levels in saliva, urine or serum. The relative proportion of EGF levels in submandibular gland saliva, parotid saliva, and whole saliva was 1:6:4. The positive rate of immunohistochemical EGF showed no significant differences between submandibular gland, parotid gland, sublingual gland or minor salivary gland. Salivary EGF levels were markedly low in patients with oral inflammations (stomatitis aphthosa, or peritonsillar abscess) or head and neck tumors (squamous cell carcinoma of the tongue, oral cavity, hypopharynx or larynx). These findings may be significant pathophysiologically. Low salivary EGF levels may reduce the capacity of oral mucosal defense mechanisms to fight against injury by physiochemical agents.

Topics: Adolescent; Adult; Aged; Child; Child, Preschool; Circadian Rhythm; Epidermal Growth Factor; Female; Head and Neck Neoplasms; Humans; Male; Middle Aged; Radioimmunoassay; Reference Values; Saliva; Salivary Gland Neoplasms; Salivary Glands; Sialadenitis; Stomatitis

Retinoic acid (RA) binding has been detected in the nuclei of a subclone (CL-1) of human submandibular adenocarcinoma cell line HSG conditioned to grow in a serum-free defined medium. Competition assay confirmed the specificity of the RA binding. Scatchard analysis showed the binding molecule to have a high affinity and low capacity. From the analyses by gel-filtration and glycerol density gradient centrifugation, the nuclear binding molecule appears to be distinct from cellular RA binding protein (CRABP) in terms of molecular weight. Furthermore, immunoblotting analysis revealed a band (Mr 47,000) reactive with specific antibody to RA receptor (RAR) alpha in the gel containing the nuclear fraction of CL-1 cells. Northern blotting analysis with specific cDNA probes revealed the expression of RAR alpha and RAR gamma in CL-1 cells. These results indicate that CL-1 cells express two types of RAR subtype, suggesting that these receptor molecules may mediate biological effects of RA. Treatment of CL-1 cells with RA resulted in an increase in the incorporation of [3H]thymidine into TCA-insoluble materials. The maximal increase was observed at 10(-6) M around 48 h. Previously, we demonstrated the autocrine growth of HSG cells mediated by epidermal growth factor (EGF) receptors and EGF-like molecules (Kurokawa et al. (1989) Cancer Res. 49, 5136-5142) and showed that RA had no significant effect on the secretion of the EGF-like molecule. RA induced an increase in [125I]EGF binding to CL-1 cells. The increase in the EGF binding was maximal at 24 h at 10(-6) M RA. RA also increased the amount of [3H]leucine-labeled EGF receptor dose-dependently. No significant change was observed in total protein synthesis of CL-1 cells by treatment with RA. These results suggest that RA stimulates the growth of CL-1 cells by increasing EGF receptor levels.

Topics: Adenocarcinoma; Carrier Proteins; Cell Division; Cell Nucleus; Clone Cells; DNA, Neoplasm; Epidermal Growth Factor; ErbB Receptors; Humans; Receptors, Retinoic Acid; RNA, Messenger; Salivary Gland Neoplasms; Tretinoin; Tumor Cells, Cultured

Dexamethasone (1 microM) decreased the distribution of cells in S phase (about 75%) and increased that of G1 cells (1.1-fold) in the DNA histogram of human submandibular salivary gland adenocarcinoma cells (HSG) reversibly. In synchronized cells at G1 phase, glucocorticoid delayed the initiation of DNA synthesis by about 3-4 h. The conditioned medium (50%) or exogenous human epidermal growth factor (EGF, 10 ng/ml) significantly nullified these effects by glucocorticoids. These results suggested that glucocorticoids arrested the cells at G1 phase, which implied the inhibition of production of some progressive factor, probably EGF, in the cell cycle of HSG.

Topics: Adenocarcinoma; Cell Division; Culture Media; Dexamethasone; DNA; Epidermal Growth Factor; G1 Phase; Humans; S Phase; Salivary Gland Neoplasms; Triamcinolone Acetonide; Tumor Cells, Cultured

A neoplastic human salivary intercalated duct cell line (HSG) and its derivatives, HSG with a myoepithelial cell phenotype (HSG-AZA1) and HSG with an acinar cell phenotype (HSG-AZA3), which were induced by 5-azacytidine treatment of HSG cells, were cultivated in the presence of epidermal growth factor (EGF). Morphological changes occurred; cells that were spindle shaped or stellate and had long cytoplasmic processes appeared in all of the treated cells. Major alterations, such as expression of neuron-specific enolase, neurofilaments, or synaptophysin as well as formation of neurite-like structures densely packed with microfibrils and microtubules, were observed in these cells with a phenotype similar to that of neuron-like cells. Both the anchorage-independent and anchorage-dependent growths of the treated HSG cells and only anchorage-dependent growth of the treated HSG-AZA1 cells were suppressed, whereas the anchorage-dependent growth of the treated HSG-AZA3 cells was enhanced according to the increasing concentrations of EGF. In addition, it has been found by statistical analysis that the growth of HSG cells in culture is suppressed and that of HSG-AZA3 cells in culture is stimulated according to the increasing concentrations of EGF added, whereas the growth of cultured HSG-AZA1 cells is hardly affected by EGF treatment. The HSG, HSG-AZA1, and HSG-AZA3 cell had 1.8 x 10(5), 1.83 x 10(5), and 1.34 x 10(5) EGF receptors, respectively. The amounts of the EGF internalized into cells were larger in HSG-AZA3 cells than in HSG or HSG-AZA1 cells. These findings indicate that the conversion of HSG, HSG-AZA1, and HSG-AZA3 cells into neuron-like cells occurs in growth medium containing EGF, with a concomitant modulation of the growth potentials of the cells which were examined in a different manner.

Topics: Amylases; Biomarkers, Tumor; Carcinoembryonic Antigen; Cell Division; Dose-Response Relationship, Drug; Epidermal Growth Factor; Humans; Immunoenzyme Techniques; In Vitro Techniques; Intermediate Filaments; Membrane Proteins; Myosins; Phosphopyruvate Hydratase; Salivary Gland Neoplasms; Synaptophysin; Tumor Cells, Cultured

Human salivary gland adenocarcinoma cell line HSG secretes an epidermal growth factor (EGF)-like molecule and contains EGF receptors. Growth of HSG cells is inhibited by glucocorticoid. We have identified that the growth inhibition by glucocorticoid is induced by the reduced secretion of the EGF-like molecule and that addition of anti-human EGF antibody to the culture specifically inhibits the growth of HSG cells, suggesting that autocrine secretion is involved in the growth of HSG cells. To prove that autocrine secretion functions in glucocorticoid-regulated growth of the HSG cell line, we purified the EGF-like molecule from serum-free, defined medium conditioned by the HSG cells and examined the growth-stimulatory effect of the purified molecule. The cultivation of HSG cells in serum-free defined medium, which contains insulin (10 micrograms/ml) and transferrin (10 micrograms/ml) only as proteinaceous components, resulted in establishment of a new cell line (HSG-SF) which had different morphological features from the parental HSG cell line. HSG-SF cells were found to have basically the same responsiveness to glucocorticoid as parental HSG cells. Parental HSG cells secreted high molecular weight EGF-like molecules (Mr 46,000 and 57,000), which were recognized by specific antibody to low molecular weight human EGF (Mr 6,201). From conditioned, serum-free medium of HSG-SF cells, an EGF-like molecule (Mr 46,000) was purified by using an anti-human EGF antibody-coupled Sepharose CL-4B column. This EGF-like molecule induced a maximal increase (36%) in incorporation of [3H]thymidine into DNA of parental HSG cells as well as low molecular weight human EGF. These observations demonstrate that growth of the HSG cell line is regulated by autocrine secretion.

Topics: Adenocarcinoma; Cell Division; Cell Line; Culture Media; DNA Replication; Epidermal Growth Factor; Fibronectins; Humans; Salivary Gland Neoplasms; Triamcinolone Acetonide; Tumor Cells, Cultured

The phenotypic expression of the human epidermal growth factor (EGF) was investigated immunohistochemically in human foetal submandibular glands from the 5th to 10th month of gestation, adult normal submandibular glands and 48 cases of pleomorphic adenomas. In foetal submandibular glands, both the terminal buds and primary ducts at the intermediate stage of gestation were positive for EGF, and in particular, the outer layer cells of primary ducts showed strong EGF-immunoreactivity. EGF-positive cells decreased as the gestational stage advanced and only ductal cells were weakly positive for EGF at the terminal stage of gestation. In the adult normal submandibular gland, weak immunoreactivity for EGF was restricted to ductal cells. However, 41 (86%) of the 48 pleomorphic adenomas had EGF-positive cells which were distributed among the ductal, chondroid and myxoid portion. No EGF-immunoreactivity was detected in the solid portion of pleomorphic adenomas. These results suggest that EGF may play an important role in the growth and differentiation of foetal cells as well as the proliferation of tumour cells in pleomorphic adenomas.

Topics: Adenoma, Pleomorphic; Antibodies; Epidermal Growth Factor; Fetus; Humans; Immunohistochemistry; Phenotype; Salivary Gland Neoplasms; Submandibular Gland Neoplasms

The treatment of a human submandibular gland adenocarcinoma cell line (HSG cell line) for 48 h with triamcinolone acetonide (TA; 1-100 nmol/l) reduced the secretion of epidermal growth factor (EGF) in a closely related manner to a maximum of 66%. The reduction in the level of EGF secreted resulted in the suppression of DNA synthesis in the HSG cells to a similar extent. When the cells were incubated with TA and exogenous human EGF (hEGF), DNA synthesis was 1.7-fold higher than that without added hEGF. The removal of EGF by the addition of hEGF antibody reduced DNA synthesis in HSG cell cultures to the same extent as did TA. These results suggest that the growth inhibition of HSG cells by TA is due to the reduction in the amount of EGF secreted.

Topics: Adenocarcinoma; Cell Line; DNA, Neoplasm; Epidermal Growth Factor; Humans; Salivary Gland Neoplasms; Submandibular Gland Neoplasms; Triamcinolone Acetonide; Tumor Cells, Cultured

Immunohistochemical identification of human epidermal growth factor (hEGF) was carried out in a total of 152 cases of salivary gland tumours, consisting 107 pleomorphic adenomas and their variants, 13 adenolymphomas and 32 adenoid cystic carcinomas. A high percentage of pleomorphic adenomas revealed markedly positive hEGF staining of the luminal surface cells of tubuloductal structures and of modified or neoplastic myoepithelial cells. Clear cells of the tumour showed various reactivities from very slight to strong. Eosinophilic epithelial cells of adenolymphoma gave a positive reaction for hEGF in all the cases, whereas most adenoid cystic adenoma lacked hEGF staining; however some cases showed positive staining of the tumour cells. The immunohistochemical detection of hEGF in most salivary gland tumours suggests this factor to be a possible new marker of salivary glands tumours, and to have a biological role in tumour proliferation.

Topics: Adenolymphoma; Adenoma, Pleomorphic; Carcinoma, Adenoid Cystic; Epidermal Growth Factor; Histocytochemistry; Humans; Salivary Gland Neoplasms

Immunohistochemical distribution of human epidermal growth factor (hEGF) was described in 17 cases of mixed tumour of the skin with monoclonal antibody. In normal sweat glands, epithelial cells in the secretory portion and in the transitional area between secretory portion and duct showed prominent staining for hEGF. In the salivary pleomorphic adenoma type of mixed tumour of the skin, luminal tumour cells of tubular and duct-like structures gave a very characteristic hEGF staining reaction. The tumour cells showing strong staining for hEGF were scattered throughout the solid foci in this type of mixed tumour. Tubular epithelial cells in the clear cell adenoma type also displayed a positive hEGF reaction. And apocrine mixed tumours strong staining for hEGF occurred on the apical side of tubular and ductal tumour cells. In view of the immunohistochemical staining patterns for hEGF, the histologic origin of mixed tumours of the skin is suggested to be cells in the secretory portion and those in the transitional portion between secretory portion and duct of the sweat gland.

Topics: Adenoma, Pleomorphic; Antibodies, Monoclonal; Epidermal Growth Factor; Humans; Immunohistochemistry; In Vitro Techniques; Neoplasms, Germ Cell and Embryonal; Salivary Gland Neoplasms; Sweat Gland Neoplasms

We applied immunohistochemical procedures to detect hEGF in salivary glands and pleomorphic adenomas of salivary-gland origin using a polyclonal hEGF antiserum and a monoclonal antibody against hEGF synthesized by applying the synthetic gene technique using Escherichia coli. In normal salivary glands, hEGF was mainly localized in the ductal system (i.e., intercalated, striated, and excretory ducts). The staining intensity and intracellular localization exhibited some variation depending on the fixative used. When a polyclonal hEGF antiserum was used for immunostaining, slight background staining was observed in sections prepared using the fixatives tested. Therefore, precise localization of hEGF was obtainable only in formalin-fixed sections using the monoclonal antibody against hEGF. In pleomorphic adenomas, positive hEGF staining was seen on the luminal side of tumors and in cells of ductal origin; no reactivity was present on the outer side of tumors or in cells of myoepithelial origin. Occasionally, long, spindle-shaped tumor cells and chondroidally changed tumor cells also exhibited positive staining for hEGF.

Topics: Adenoma; Antibodies, Monoclonal; Epidermal Growth Factor; Humans; Immune Sera; Immunodiffusion; Immunohistochemistry; Radioimmunoassay; Salivary Gland Neoplasms; Submandibular Gland; Submandibular Gland Neoplasms

Neoplastic intercalated duct cells that originated from a human submandibular salivary gland release transforming growth factor and human epidermal growth factor into serum-free medium. The transforming growth factor with the soft agar colony-forming activity when assayed in the presence of mouse epidermal growth factor on normal rat kidney clone 49F indicator cells, but without mitogenic action to quiescent 3T3 cells, does not compete with epidermal growth factor for receptor binding, is heat and acid resistant but trypsin and dithiothreitol sensitive, and therefore is of the transforming growth factor-beta class. The immunoreactive human epidermal growth factor is detected by radioimmunoassay on certain fractions obtained by the Bio-Gel P-60 chromatography of neoplastic epithelial duct cells originated from a human submandibular salivary gland-conditioned medium and is biologically very similar to mouse epidermal growth factor. Moreover, the presence of human epidermal growth factor in cultured neoplastic epithelial duct cells originated from a human submandibular salivary gland is demonstrated by the peroxidase:antiperoxidase method.

Topics: Adenocarcinoma; Cell Division; Cell Line; Culture Media; Epidermal Growth Factor; ErbB Receptors; Growth Substances; Humans; Peptide Biosynthesis; Peptides; Receptors, Cell Surface; Salivary Gland Neoplasms; Transforming Growth Factors

Immunohistochemical identification of epidermal growth factor (EGF) is described in experimental carcinoma of the submandibular gland of mice given testosterone before sacrifice. EGF in the submandibular gland was confined to the granular convoluted tubule (GCT) cells, and its level was enhanced following testosterone injection. In the initial phase of carcinogenesis of the gland, degranulation of the GCT cells occurred as well as decreased EGF staining in the degranulated cells. In the testosterone-treated animals, changed GCT cells showed intense EGF deposition. Histological aspects during carcinogenesis in submandibular glands indicated duct-like structures, squamous metaplasia, and squamous cell types of carcinoma with different keratinization. Immunohistochemically detectable EGF was characterized by positive staining in pre-neoplastic or early neoplastic epithelial structures in testosterone-treated mice. However, tumour epithelia did not show any EGF reaction.

Topics: Animals; Carcinoma, Squamous Cell; Epidermal Growth Factor; Female; Histocytochemistry; Male; Mice; Mice, Inbred Strains; Salivary Gland Neoplasms; Submandibular Gland Neoplasms; Testosterone

Immunohistochemical demonstration of epidermal growth factor (EGF) and nerve growth factor (NGF) was made during chemical carcinogenesis in the mouse submandibular gland. The granular convoluted tubule cells in the normal male submandibular gland contained larger amounts of EGF and NGF than in the female. The initial phase and early stages in chemical carcinogenesis showed degranulation of the granular convoluted tubule cells with a marked decrease in EGF and NGF. Premalignant lesions such as duct-like structures and multicystic lesions showed variable staining for EGF and were usually negative for NGF. Material secreted into the luminal spaces revealed increased staining for EGF and NGF. Scattered tumor cells of the poorly differentiated squamous-cell carcinoma type and desquamated tumor cells contained abundant EGF, but not NGF. No positive reaction for EGF or NGF was found in the induced squamous-cell carcinoma cells.

Topics: Animals; Carcinoma, Squamous Cell; Cell Transformation, Neoplastic; Cytoplasmic Granules; Epidermal Growth Factor; Female; Male; Mice; Nerve Growth Factors; Salivary Gland Neoplasms; Submandibular Gland Neoplasms

Dimethylbenz[a]anthracene was injected into the submandibular glands of male Swiss-Webster mice. From tumors obtained, three cell lines were established. Immunocytochemical stainings revealed epidermal growth factor, renin, and peptidase in a significant portion of cultured tumor cells. In addition, the presence of epidermal growth factor was demonstrated by radioimmunoassays. Since epidermal growth factor, renin, and peptidase are localized in the granular convoluted tubules in mouse submandibular gland, the data suggest that the granular convoluted tubule cells are the targets of chemical carcinogens.

Topics: Animals; Cell Line; Endopeptidases; Epidermal Growth Factor; Male; Methods; Peptides; Radioimmunoassay; Rats; Renin; Salivary Gland Neoplasms; Submandibular Gland