transforming-growth-factor-beta and Salivary-Gland-Neoplasms

Recently, histologic grade was removed from salivary tumor nomenclature by the WHO to include disease of higher grade. One such entity, cribriform adenocarcinoma (CAC), is an aggressive group of polymorphous adenocarcinoma (PAC), with frequent nodal metastasis and locoregional recurrence. We aim to examine the biologic behavior of this disease as compared with the PAC general cohort inclusive of all subtypes.. A systematic review of the literature on polymorphous adenocarcinoma and cribriform adenocarcinoma was completed. A descriptive analysis was performed for the following predictor variables: nodal and distant metastasis, in addition to recurrence. The outcome variables, disease free recurrence, and disease specific survival, where plotted using Kaplan-Meier curves.. PAC and CAC both show median age of diagnosis in the sixth decade of life and a female predominance. CAC occurs most frequently in the tongue and PAC in the palate. The 2 groups show a similar biologic behavior in regards to incidence of distant metastasis (4.1 vs 5.5%), recurrence (12.5 vs 17.8%), and death from disease (3 vs 2.7%). However, there was an increased incidence of nodal metastasis in CAC (53%) as compared with that in PAC of all subtypes (14%).. CAC exhibits more aggressive biologic behavior as compared with the PAC cohort. Although CAC is not an officially recognized entity, these tumors likely comprise a significant portion of the cases of PAC with poor outcomes and are deserving of attention and consideration for escalation in oncologic treatment.

Topics: Adenocarcinoma; Aggression; Female; Humans; Medical Oncology; Neoplasm Recurrence, Local; Salivary Gland Neoplasms; Transforming Growth Factor beta

Topics: Animals; Apoptosis; bcl-2-Associated X Protein; Carcinoma, Squamous Cell; Cell Line, Tumor; Induced Pluripotent Stem Cells; Male; Rats; Rats, Wistar; RNA, Long Noncoding; RNA, Neoplasm; Salivary Gland Neoplasms; Sirtuin 1; Submandibular Gland; Transforming Growth Factor beta

PLAG1 rearrangements have been described as a molecular hallmark of salivary gland pleomorphic adenoma (PA), carcinoma ex pleomorphic adenoma (CEPA), and myoepithelial carcinoma (MECA). Several fusion partners have been described, however, commonly no further assignment to the aforementioned entities or a morphological prediction can be made based on the knowledge of the fusion partner alone. In contrast, TGFBR3-PLAG1 fusion has been specifically described and characterized as an oncogenic driver in MECA, and less common in MECA ex PA. Here, we describe the clinicopathological features of three TGFBR3-PLAG1 fusion-positive salivary gland neoplasms, all of which arose in the deep lobe of the parotid gland. Histopathology showed high morphological similarities, encompassing encapsulation, a polylobular growth pattern, bland basaloid and oncocytoid cells with myoepithelial differentiation, and a distinct sclerotic background. All cases showed at least limited, unusual foci of minimal invasion into adjacent salivary gland tissue, including one case with ERBB2 (Her2/neu) amplified, TP53 mutated high-grade transformation, and lymph node metastases. Of note, all cases illustrated focal ductal differentiation. Classification remains difficult, as morphological overlaps between myoepithelial-rich cellular PA, myoepithelioma, and MECA were observed. However, evidence of minimal invasion advocates classification as low-grade MECA. This case series further characterizes the spectrum of uncommon cellular myoepithelial neoplasms harboring TGFBR3-PLAG1 fusion, which show recurrent minimal invasion of the adjacent salivary gland tissue, a predilection to the deep lobe of the parotid gland, and potential high-grade transformation.

Topics: Adenoma, Pleomorphic; Adult; Aged; DNA-Binding Proteins; Gene Rearrangement; Humans; Male; Neoplasm Grading; Oncogene Proteins, Fusion; Parotid Gland; Proteoglycans; Receptors, Transforming Growth Factor beta; Salivary Gland Neoplasms; Transforming Growth Factor beta

Tumor necrosis factor (TNF) receptor-associated factor 6 (TRAF6) has been proved to play an important role in tumorigenesis, invasion, and metastasis. However, its precise role salivary adenoid cystic carcinoma (SACC) has not been determined. The aim of this study was to explore the role of TRAF6 in SACC including invasion and metastasis of SACC cells.. Immunohistochemistry and quantitative real-time PCR were performed in SACC tissues paired with their adjacent normal tissues to analyze the expression of TRAF6. Downstream proteins expression was explored when TRAF6 was knockdown by siRNA.. The results show that TRAF6 is upregulated in SACC samples, especially in SACC with metastasis, which is closely correlated with an aggressive phenotype (P = .0073) and shorter life survival span (P = .0061) in SACC patients. Knockdown of TRAF6 can attenuate the promotion effect of SACC cell invasion induced by TGF-β. Western blot results also showed that silencing TRAF6 expression can inhibit the activation of SMAD2, SMAD3, ERK, p38, and JNK induced by TGF-β in SACC cells.. These data suggested that TRAF6 regulates TGF-β-mediated SACC progression through SMAD2/3-ERK-p38-JNK cascades.

Topics: Carcinogenesis; Carcinoma, Adenoid Cystic; Cell Line, Tumor; Disease Progression; Female; Humans; Intracellular Signaling Peptides and Proteins; Male; MAP Kinase Signaling System; Middle Aged; Neoplasm Invasiveness; p38 Mitogen-Activated Protein Kinases; Salivary Gland Neoplasms; Smad2 Protein; Smad3 Protein; TNF Receptor-Associated Factor 6; Transforming Growth Factor beta; Up-Regulation

Salivary gland tumor (SGT) is a rare tumor type, which exhibits broad-spectrum phenotypic, biological, and clinical heterogeneity. Currently, the molecular mechanisms that cause SGT pathogenesis remain poorly understood. A lack of animal models that faithfully recapitulate the naturally occurring process of human SGTs has hampered research progress on this field. In this report, we developed an inducible keratin 5-driven conditional knockout mouse model to delete gene(s) of interest in murine salivary gland upon local RU486 delivery. We have deleted two major tumor suppressors, Pten, a negative regulator of the PI3K pathway, and Smad4, the central signaling mediator of TGFβ pathway, in the murine salivary gland. Our results have shown that deletion of either Pten or Smad4 in murine salivary gland resulted in pleomorphic adenomas, the most common tumor in human SGT patients. Deletion of both Pten and Smad4 in murine salivary gland developed several malignancies, with salivary adenoid cystic carcinoma (SACC) being the most frequently seen. Molecular characterization showed that SACC exhibited mTOR activation and TGFβ1 overexpression. Examination of human SGT clinical samples revealed that loss of Pten and Smad4 is common in human SACC samples, particularly in the most aggressive solid form, and is correlated with survival of SACC patients, highlighting the human relevance of the murine models. In summary, our results offer significant insight into synergistic role of Pten and Smad4 in SGT, providing a rationale for targeting mTOR and/or TGFβ signaling to control SGT formation and progression.

Topics: Animals; Carcinoma, Adenoid Cystic; Disease Progression; Humans; Mice; Mice, Inbred C57BL; Mifepristone; Phosphatidylinositol 3-Kinases; PTEN Phosphohydrolase; Salivary Gland Neoplasms; Salivary Glands; Signal Transduction; Smad4 Protein; TOR Serine-Threonine Kinases; Transforming Growth Factor beta

Epithelial-mesenchymal transition (EMT) plays an important role in the invasion and metastasis of salivary adenoid cystic carcinoma (SACC) which is characterized by wide local infiltration, perineural spread, a propensity to local recurrence and late distant metastasis. Our recent studies have disclosed that TGF-β is a crucial factor for EMT in metastatic SACC. In this study, we further uncovered small redox protein thioredoxin 1 (TXN) as a critical mediator of TGF-β induced EMT. Immunohistochemistry analysis revealed significantly higher expressions of TXN, thioredoxin reductase 1 (TXNRD1) and N-cadherin, and lower expression of E-cadherin in human metastatic SACC compared to non-metastatic SACC tissues. Consistently, cultured SACC cells with stable TXN overexpression had decreased E-cadherin and increased N-cadherin as well as Snail and Slug expressions. The enhanced migration and invasion potential of these cells was abrogated by Akt or TXNRD1 inhibitors. Expression of N-cadherin and Akt p-Akt decreased, whereas E-cadherin expression increased in a BBSKE (TXNRD1 inhibitor)-dose-dependent manner. In a xenograft mouse model, TXN overexpression facilitated the metastatic potential of SACC-83 cells to the lung. Our results indicate that TXN plays a key role in SACC invasion and metastasis through the modulation of TGF-β-Akt/GSK-3β on EMT. TXN could be a potential therapeutic target for SACC.

Topics: Adult; Animals; Antigens, CD; Cadherins; Carcinoma, Adenoid Cystic; Cell Line, Tumor; Cell Movement; Epithelial-Mesenchymal Transition; Female; Gene Expression Regulation, Neoplastic; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Humans; Kaplan-Meier Estimate; Lung Neoplasms; Male; Mice, Inbred NOD; Mice, SCID; Middle Aged; Neoplasm Invasiveness; Phosphorylation; Proto-Oncogene Proteins c-akt; RNA Interference; Salivary Gland Neoplasms; Signal Transduction; Snail Family Transcription Factors; Thioredoxin Reductase 1; Thioredoxins; Time Factors; Transcription Factors; Transfection; Transforming Growth Factor beta

Epithelial-mesenchymal transition (EMT) is associated with salivary adenoid cystic cancer (ACC) progression and metastasis. Here, we report that ectopic overexpression of c-kit in ACC cell lines is sufficient for acquisition of mesenchymal traits, enhanced cell invasion, along with stem cell properties defined by the presence of a CD133+/CD44+ cell subpopulation. c-kit positively regulated expression of known EMT inducers, also activating TGF-β to contribute to EMT. c-kit itself was induced by TGF-β in ACC cell lines and required for TGF-β-induced EMT. Xenograft experiments showed that c-kit cooperated with oncogenic Ras to promote tumorigenesis in vivo. Finally, in human specimens of ACC, we found that c-kit was abnormally overexpressed and correlated with the prognosis of ACC. Our findings define an important function for c-kit in ACC progression by orchestrating EMT, and they implicate this gene product as a marker of poor prognosis in this disease.

Topics: Apoptosis; Blotting, Western; Carcinoma, Adenoid Cystic; Cell Movement; Cell Proliferation; Cell Transformation, Neoplastic; Disease Progression; Epithelial-Mesenchymal Transition; Flow Cytometry; Fluorescent Antibody Technique; Gene Expression Regulation, Neoplastic; Genes, ras; Humans; Immunoenzyme Techniques; Neoplasm Invasiveness; Neoplastic Stem Cells; Prognosis; Proto-Oncogene Proteins c-kit; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Salivary Gland Neoplasms; Signal Transduction; Survival Rate; Transforming Growth Factor beta; Tumor Cells, Cultured

Adenoid cystic carcinoma (ACC) is one of the most common malignancies of salivary glands, characterized by poor prognosis, particularly due to pulmonary metastasis. We previously reported that transforming growth factor (TGF)-β1 promoted ACC cell migration and invasion via the Smad pathway in vitro. The aim of this study was to establish the underlying mechanisms.. TGF-β1, phospho-Smad2 and β-catenin expression in ACC tissues derived from patients was evaluated by immunohistochemistry. The role of TGF- β1 on the invasive capacity of ACC cells was determined by transwell assays in SACC-83 cells transfected with TGF-β1 and TGF-β type II dominant-negative receptor (TβRIIDN) plasmids or silenced by TGF-β1 siRNA. Expression of the epithelial-mesenchymal transition (EMT) markers, β-catenin, E-cadherin and Nectin-1, was determined by real-time PCR and immunochemistry. In vivo investigations were performed by inoculating nude mice with the transfected ACC cells and examining metastasis in bilateral lung tissues by immunohistochemistry.. Overexpression of TGF-β1 and phospho-Smad2, and reduced expression of membrane β-catenin, were closely associated with lung metastasis in ACC. Furthermore, the EMT markers were downregulated. In vitro, cells transfected with TGF-β1 exhibited altered morphology and increased invasive capacity compared to TβRIIDN-transfected cells or TGF-β1 siRNA silenced cells. In vivo, mice inoculated with TGF-β1 transfected ACC cells exhibited more metastases than other cells.. TGF-β1, phospho-Smad2 and β-catenin were significantly correlated with ACC metastasis. Blockade of TGF-β signaling by TβRIIDN or siRNA may offer potential gene therapies against pulmonary metastasis in patients with ACC.

Topics: Animals; beta Catenin; Cadherins; Carcinoma, Adenoid Cystic; Cell Adhesion Molecules; Epithelial-Mesenchymal Transition; Female; Humans; Lung Neoplasms; Male; Mice; Mice, Nude; Middle Aged; Nectins; Neoplasms, Experimental; Real-Time Polymerase Chain Reaction; Salivary Gland Neoplasms; Smad2 Protein; Transforming Growth Factor beta

Mesenchymal stem cells (MSCs) are the progenitors of stromal cells, which have been found to interact with cancer cells and represent an important target for cancer therapies. Salivary gland cancer is an aggressive malignant epithelial tumor and can easily disseminate to distant sites in vivo. In this work, we probed the role of MSCs in salivary gland cancer in an in vivo like tumor microenvironment by using a series of functional microfluidic devices for 2D and 3D assays. The results demonstrated that MSCs could be recruited by salivary gland cancer cells (ACC-M) and this effect was potentially mediated by TGF-β secreted by cancer cells. In addition, MSCs exhibited the ability to squeeze into ACC-M spheroids by dispersing the cell-cell connection and reducing the expression of E-cadherin in cancer cells. In particular, MSCs exhibited the ability to enhance the invasion of salivary cancer under a chemokine CXCL12 gradient, indicating the involvement of a CXCL12-CXCR4 pathway and the tumor-promoting properties of MSCs in cancer progression. This study recreated an in vivo-like tumor microenvironment by constructing a series of functional microdevices and explored the role of MSCs in ACC invasion for the first time. It provides a unique platform to open up new options to target stromal cells for cancer therapy and also analyzed the potential risk of using MSCs as drug delivery carriers for therapeutic purposes in carcinoma treatment.

Topics: Biomimetics; Cell Communication; Cell Line; Chemokine CXCL12; Equipment Design; Humans; Mesenchymal Stem Cells; Microfluidic Analytical Techniques; Receptors, CXCR4; Salivary Gland Neoplasms; Transforming Growth Factor beta

Salivary gland tumors (SGTs) are known for their specific heterogeneity and ambiguous outcome for the affected patients. The LKB1 and SMAD4 genes are pivotal components of important signaling pathways, including AMPK and TGF-β. To our knowledge, no study has reported an association between the expression levels of these genes in SGTs. The expression levels of LKB1 and SMAD4 were evaluated to clarify their possible crosstalk in SGTs.. A total of 50 fresh tissue specimens were obtained from patients with SGTs, including pleomorphic adenoma (PA), Warthin's tumor (WT), intermediate grade mucoepidermoid carcinoma (MEC), salivary duct carcinoma (SDC), and carcinoma ex pleomorphic adenoma (CexPA), as well as 8 normal samples. Quantitative real-time polymerase chain reaction was performed for all samples with specific primers.. Data were analyzed using statistical methods. PA, WT, MEC, and SDC showed a significant decrease in LKB1 levels, but the gene was up-regulated in CexPA. SMAD4 was overexpressed in all samples.. The results suggest a possible link between downregulation of LKB1 and overexpression of SMAD4 in SGTs. LKB1 depletion leads to upregulation of SMAD4, promoting epithelial-mesenchymal transition in tumor cells. Therefore, LKB1 and SMAD4 could be key players in inducing tumor heterogeneity in SGTs.

Topics: AMP-Activated Protein Kinase Kinases; Humans; Protein Kinases; Protein Serine-Threonine Kinases; Receptor Cross-Talk; Salivary Gland Neoplasms; Signal Transduction; Smad4 Protein; Tissue Distribution; Transforming Growth Factor beta

Radiation-induced xerostomia still represents a common symptom following radiotherapy of head and neck malignancies, which significantly impairs the patient's quality of life. In this cross-sectional study, human salivary glands were investigated to assess the role of Wnt/β-catenin and TGF-β pathways in the pathogenic process of radiogenic impairment of salivary function.. Irradiated human salivary glands were investigated in patients with manifested xerostomia. Alteration of Wnt-1 and cell-cell adhesion was evaluated immunohistologically as well as changes in the expression of TGF-β were assessed in salivary gland tissue.. We assessed two alteration patterns in which Wnt-1 expression represents one change along with up-regulation of β-catenin and E-cadherin in irradiated but viable acinar cells. Increased expression of tenascin-C was observed in sites of epithelial-mesenchymal interaction and loss of cell-cell adhesion was assessed in translocated epithelial cells in the stroma.. Increased transdifferentiation and remodeling of acinar structures was associated with decrease of viable acinar structures. The role of Wnt and TGF signaling may provide a potential therapeutic approach to prevent radiation-induced damage to salivary glands during radiotherapy for head and neck cancer.

Topics: Cadherins; Cell Adhesion Molecules; Cross-Sectional Studies; Female; Fluorescent Antibody Technique; Gene Expression Regulation, Neoplastic; Head and Neck Neoplasms; Humans; Immunohistochemistry; Male; Prognosis; Radiotherapy; Radiotherapy Dosage; Risk Assessment; Salivary Gland Neoplasms; Salivary Glands; Transforming Growth Factor beta; Treatment Outcome; Up-Regulation; Wnt1 Protein; Xerostomia

Defects in the mitotic checkpoint lead to aneuploidy and might facilitate tumorigenesis. However, the ploidy status in salivary duct carcinoma (SDC) has been reported to play limited role in prediction of prognosis. Thus, we need more reliable markers to reflect the rapid tumor progression in SDCs. We aimed here to investigate the expression of mitotic checkpoint proteins benzimidazole 1 homolog beta (BUB1B) and mitosis arrest-deficient 2 like 1 (MAD2L1) in SDCs and to determine their possible role as surrogate prognostic markers.. We analyzed the clinical courses, pathologic findings and immunohistochemical profiles of mitotic checkpoint proteins (BUB1B and MAD2L1) in 27 pathologically confirmed SDCs. The expression status of BUB1B and MAD2L1 was compared with clinicopathologic factors and other molecular markers, such as TGF-beta, c-erb-B2, androgen receptor, vascular endothelial growth factor, and epidermal growth factor receptor, for prognostic significance.. High BUB1B expression was detected in 25.9% of subjects, and high MAD2L1 expression was in 55.6% of subjects. However, survival analysis revealed that mitotic checkpoint expression did not have prognostic significance in SDCs, nor did the other studied markers. Rather, the clinical variable of N classification at diagnosis (in N+ status, hazard ratio 5.19, 95% CI 1.26-21.32 for disease-free survival and hazard ratio 7.18, 95% CI 1.09-46.99 for overall survival) was strongly associated with survival and prognosis based on the Cox proportional hazard model.. Mitotic checkpoint proteins appeared to play a limited role in predicting prognosis in SDCs. Further study is required to elucidate the exact role of mitotic checkpoint proteins in SDCs.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Calcium-Binding Proteins; Carcinoma; Cell Cycle Proteins; Cohort Studies; Disease-Free Survival; ErbB Receptors; Female; Gene Expression Regulation, Neoplastic; Humans; Mad2 Proteins; Male; Middle Aged; Neoadjuvant Therapy; Neoplasm Staging; Proportional Hazards Models; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins; Receptor, ErbB-2; Receptors, Androgen; Remission Induction; Repressor Proteins; Salivary Ducts; Salivary Gland Neoplasms; Survival Rate; Transforming Growth Factor beta; Treatment Outcome; Vascular Endothelial Growth Factor A

Runt-related transcription factor-3 (RUNX3), being a tumor suppressor gene in gastric cancer, plays an important role in inhibiting cellular growth by participating in the transforming growth factor-beta-dependent apoptosis. The aim of this study was to determine the expression of RUNX3 in normal salivary glands and adenoid cystic carcinomas (ACCs), comparing the results with clinicopathological factors and patient survival. The quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis and Western blot analysis revealed the expression of RUNX3 both in normal salivary glands and ACCs. Nuclear and cytoplasmic immunoreactivities against RUNX3 in ductal luminal cells and acinous cells, but immunonegative in myoepithelial cells, were detected in normal salivary glands. In ACC, the RUNX3 immunostaining was shown in the cytoplasm of tumor cells; however, no nuclear location of RUNX3 was found. Lower RUNX3 expression showed significant correlation to distant metastasis and histological growth pattern (P = 0.009 and P = 0.025, respectively). On univariate analysis, low level of RUNX3 immunolabeling (P = 0.012), stage T4 (P = 0.017), lymph node involvement (P = 0.007), and distant metastasis (P < 0.001) were significantly associated with decreased overall survival. Multivariate analysis showed only distant metastasis had an independent prognostic effect on overall survival (P = 0.043). Our results demonstrate the expression of RUNX3 in normal salivary glands and salivary ACCs. The low level of RUNX3 protein in salivary ACCs might play a pivotal role in tumor progression and have prognostic values in ACCs.

Topics: Blotting, Western; Carcinoma, Adenoid Cystic; Core Binding Factor Alpha 3 Subunit; Cytoplasm; Disease Progression; Humans; Immunohistochemistry; Middle Aged; Prognosis; Reverse Transcriptase Polymerase Chain Reaction; Salivary Gland Neoplasms; Salivary Glands; Transforming Growth Factor beta

Mucoepidermoid carcinoma (MEC) is the most common malignant salivary tumour, classified as low, intermediate and high grade. Myofibroblasts are the main stromal component and are included as prognostic factor in some tumours. The aim of this study was to evaluate the myofibroblasts in the stroma of MEC with possible relationship to malignancy grading.. Twenty-five cases of MEC (six low grade, 11 intermediate grade, four high grade and four metastasis) were stained for vimentin, desmin and smooth muscle actin (SMA) for the identification of myofibroblasts. Transforming growth factors (TGFbeta1 and TGFbetaRII) were also assessed in our study.. Myofibroblasts were present in all cases, in amounts varying according to histological grading. TGFbeta1 was positive in squamous cells of intermediate grade tumours, and in the stroma of only four cases. TGFbetaRII was positive in most squamous and intermediate cells, regardless of malignancy grading.. Our study showed that the analysis of neoplastic stroma must be added to the studies of neoplastic cells to draw a better picture leading to tumour diagnosis and prognosis.

Topics: Actins; Carcinoma, Mucoepidermoid; Desmin; Fibroblasts; Humans; Immunohistochemistry; Neoplasm Proteins; Prognosis; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type II; Receptors, Transforming Growth Factor beta; Salivary Gland Neoplasms; Stromal Cells; Transforming Growth Factor beta; Transforming Growth Factor beta1; Vimentin

To determine the possible role of TGF beta 1 and TGF beta RI in facilitating neoplastic progression in human salivary adenoid cystic carcinoma (ACC).. The expression of TGF beta 1 and TGF beta RI protein in human salivary adenoid cystic carcinoma was detected by the streptavidin-peroxidase (S-P) immunohistochemical method, and their contents were compared with those of normal salivary gland tissues and salivary pleomorphic adenoma (PA).. 1. TGF beta 1 and TGF beta RI proteins were expressed in most ductal epithelial cells and myo-epithelial cells, but they were not expressed in the acinar epithelial cells of any normal glands examined; 2. TGF beta 1 and TGF beta RI proteins were expressed in the duct-like arranging tumor cells and other solid proliferating tumor cells of salivary pleomorphic adenoma, and no difference of their contents existed between PA and normal salivary gland tissues (P > 0.05); 3. The contents of TGF beta 1 in ACC were significantly higher than those in normal salivary glands (P < 0.01), while the contents of TGF beta RI in ACC were significantly lower than those in normal salivary glands (P < 0.01).. The abnormal expression of TGF beta 1 and TGF beta RI might play an important role in tumor genesis and the development of malignant salivary gland tumors such as ACC; the over-expression of TGF beta 1 and low-expression of TGF beta RI might involve the bad biological behavior of ACC.

Topics: Biomarkers, Tumor; Carcinoma, Adenoid Cystic; Humans; Receptors, Transforming Growth Factor beta; Salivary Gland Neoplasms; Transforming Growth Factor beta; Transforming Growth Factor beta1

Although pleomorphic adenoma is the most common type of salivary gland epithelial tumor, it frequently contains "mesenchymal"-like components, including myxoid or chondroid tissues. We reported previously that chondroid tissue formation in pleomorphic adenoma was associated with overexpression of bone morphogenetic proteins (BMPs) by neoplastic myoepithelial cells. BMPs belong to the transforming growth factor (TGF)-beta superfamily, so we hypothesized that pleomorphic adenoma may express TGF-betas and that these molecules may regulate mesenchymal-like tissue formation. To evaluate this hypothesis, we immunohistochemically examined TGF-beta1, -beta2 and -beta3 expression and localization in normal salivary glands and in 43 cases of pleomorphic adenomas. There was no evidence of TGF-beta1 expression in normal salivary glands or pleomorphic adenomas. Signals for TGF-beta2 in the normal salivary glands were observed in the intercalated ducts, while in pleomorphic adenomas they were observed in the inner ductal cells of the tubulo-glandular structures. Signals for TGF-beta3 in the normal salivary glands were observed in mucous cells, whereas in pleomorphic adenomas they were observed in the solid nests of neoplastic myoepithelial cells, in the portion showing squamous metaplasia, and in the inner ductal cells of tubulo-glandular structures. TGF-betas induce ectopic cartilage formation in vivo, but chondroid tissues in pleomorphic adenomas showed only weak TGF-beta3 expression. TGF-beta may be related to differentiation of the inner ductal cells and the neoplastic myoepithelial cells. In conclusion, pleomorphic adenomas expressed TGF-beta2 and -beta3, which may be associated with differentiation of the inner ductal cells and neoplastic myoepithelial cells.

Topics: Adenoma, Pleomorphic; Antibodies; Bone Morphogenetic Proteins; Cartilage; Cell Differentiation; Coloring Agents; Epithelial Cells; Humans; Immunoenzyme Techniques; Immunohistochemistry; Mesoderm; Metaplasia; Mucous Membrane; Salivary Ducts; Salivary Gland Neoplasms; Salivary Glands; Transforming Growth Factor beta; Transforming Growth Factor beta1; Transforming Growth Factor beta2; Transforming Growth Factor beta3

One important pathological character of pleomorphic adenomas is that there is chorndriod component in the epithelial tumors. It is well known that bone morphogenetic protein (BMPs) plays important roles in the morphogenesis of hard tissue by inducing mesenchymal cells to differentiate into osteoblasts and chondroblasts in vivo. So we examined the expression of BMP and BMP2 mRNA in pleomorphic adenomas to study the pathological mechanism of chorndriod tissue formation.. Using ABC immunohistochemistry method, 6 specimens of pleomorphic adenomas were examined with BMP antibody. Reverse transcriptase-polymerase chain reaction (RT-PCR) methods were developed for detecting BMP2 mRNA expression with special primers used to amplify the BMP2 cDNA mature domain fragment. The total RNA was extracted from frozen 6 specimens of pleomorphic adenoma and then reversed to cDNA by reverse transcription using AMV reverse transcriptase. PCR conditions were 94 degrees C for 20 seconds, 60 degrees C for 30 seconds, and 72 degrees C for 1 minute, 40 cycles, followed by extension for 30 minute at 72 degrees C. The reaction products were analyzed by electrophoresis.. BMP expression could be found in all of the specimens of pleomorphic adenoma examined by immunohistochemistry method. BMP2 RT-PCR product was detected in only 5 specimens that showed BMP2 mRNA existed in 5 specimens.. These results may be helpful to study the mechanism of pleomorphic adenoma. It is speculated that BMP secreted by tumor cells may play important roles in origin of osteoid tissue in pleomorphic adenoma by inducing neoplastic myoepithelial cells to differentiate to chondrocytes.

Topics: Adenoma, Pleomorphic; Bone Morphogenetic Protein 2; Bone Morphogenetic Proteins; Humans; Immunohistochemistry; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Salivary Gland Neoplasms; Transforming Growth Factor beta

We have recently isolated TSC-22 (transforming growth factor-beta-stimulated clone-22) cDNA as an anticancer, drug-inducible (with vesnarinone) gene in a human salivary gland cancer cell line, TYS. We have also reported that TSC-22 negatively regulates the growth of TYS cells and that down-regulation of TSC-22 in TYS cells plays a major role in salivary gland tumorigenesis (Nakashiro et al, 1998). In this study, we transfected TYS cells with an expression vector encoding the TSC-22-GFP (green fluorescent protein) fusion protein, and we established TSC-22-GFP-expressing TYS cell clones. Next, we examined (a) the subcellular localization of the fusion protein, (b) the sensitivity of the transfectants to several anticancer drugs (5-fluorouracil, cis-diaminedichloroplatinum, peplomycin), and (c) induction of apoptotic cell death in the transfectants by 5-fluorouracil treatment. The TSC-22-GFP fusion protein was clearly localized to the cytoplasm, but not to the nucleus. Over-expression of the TSC-22-GFP fusion protein did not affect cell growth, but significantly increased the sensitivity of the cells to the anticancer drugs (p < 0.01; one-way ANOVA). Furthermore, over-expression of the TSC-22-GFP fusion protein markedly enhanced 5-fluorouracil-induced apoptosis. These findings suggest that over-expression of TSC-22-GFP protein in TYS cells enhances the chemosensitivity of the cells via induction of apoptosis.

Topics: Antineoplastic Agents; Apoptosis; Cell Adhesion; Cell Division; Cell Survival; Chromatin; Cisplatin; Fluorouracil; Green Fluorescent Proteins; Humans; Luminescent Proteins; Peplomycin; Recombinant Fusion Proteins; Salivary Gland Neoplasms; Transcription, Genetic; Transfection; Transforming Growth Factor beta; Tumor Cells, Cultured

This study investigated the immunolocalization of small and large proteoglycans (PGs), including decorin, biglycan, PG-M/versican and aggrecan, in salivary pleomorphic adenoma (PA) using monoclonal and polyclonal antibodies. In addition, a polyclonal antibody, A0082, recognizing blood vessels was also used to help identify truly mesenchymal tissues in PA. Decorin reactivity was detected only in tumor capsule and interstitial tissue of non-neoplastic salivary gland, but not in the tumor tissue. Biglycan was frequently revealed throughout the matrix of small chondroid regions and in the peripheral portion of larger chondroid regions. PG-M/versican was mainly localized to the truly mesenchymal tissues in PA and the innermost portion of tumor capsule. On the contrary, aggrecan was extensively expressed in the non-luminal epithelial areas as well as in the myxoid and chondroid areas, but not in the truly mesenchymal tissues. These findings suggest that aggrecan is the most widely distributed PG in PA and may be produced mainly by non-luminal tumor cells. The absence of aggrecan from the truly mesenchymal tissues argues against its origin from this source. Both aggrecan and biglycan may play important roles in the chondroid differentiation and morphogenesis of PA.

Topics: Adenoma, Pleomorphic; Aggrecans; Antibodies; Antibodies, Monoclonal; Biglycan; Cell Differentiation; Chondroitin Sulfate Proteoglycans; Decorin; Epithelium; Extracellular Matrix; Extracellular Matrix Proteins; Humans; Immunohistochemistry; Lectins; Lectins, C-Type; Mesoderm; Morphogenesis; Proteoglycans; Salivary Gland Neoplasms; Salivary Glands; Transforming Growth Factor beta; Versicans

To explore the effect of expression of epidermal growth factor receptor (EGFR), transforming growth factor beta (TGF beta), c-erbB-2 oncogene in myoepithelioma and malignant myoepithelioma.. 20 cases of myoepithelioma and 20 cases of malignant myoepithelioma were examined by immunohistochemical methods.. All cases showed overexpression of EGFR and TGF beta, the intensity of expression didn't associate with the benignity and malignity of tumors. The overexpression of c-erbB-2 oncoprotein was seen in 5 cases of myoepithelioma and 13 cases of malignant myoepithelioma, and the cases of malignant myoepithelioma were stained more intense than that of myoepithelioma.. The synergy of EGFR and TGFR beta had significant relation to the proliferative and differentiation processes of myoepithelial tumor. The activated c-erbB-2 oncogene, that is similar with EGFR in structure, plays an important promoting role in unrestricted progression of malignant myoepithelioma cells.

Topics: Biomarkers, Tumor; ErbB Receptors; Humans; Immunohistochemistry; Myoepithelioma; Receptor, ErbB-2; Salivary Gland Neoplasms; Transforming Growth Factor beta

Immunohistochemical investigation of bone morphogenetic protein-2 (BMP-2) and type II collagen, two cartilage-associated proteins, was undertaken using monoclonal antibodies in 20 cases of salivary pleomorphic adenoma (PA) in order to explore their possible roles in chondroid differentiation of this tumor. Other salivary gland tumors, including adenoid cystic carcinoma (17 cases), polymorphous low-grade adenocarcinoma (10 cases), basal cell adenoma (3 cases), basal cell adenocarcinoma (1 case), and epithelial-myoepithelial carcinoma (2 cases), were also examined for comparison. In PA, BMP-2 immunoreactivity was detected in the luminal and non-luminal cells of the tubulo-ductal structures, plasmacytoid cells, and other scattered tumor cells in solid areas. In addition, tumor cells in chondroid areas in most cases (14/15), and stellate cells in myxoid areas in many cases (7/19), were also intensely labeled for BMP-2. Furthermore, BMP-2 was also detected in the non-neoplastic ductal cells in salivary glands, whereas no other salivary gland tumors were positively stained for this protein. Type II collagen was localized in the intercellular matrix of chondroid areas and in a few chondroid differentiating cells in myxoid areas, confirming its cartilage-specificity. A proportional relationship was observed between BMP-2 expression and chondroid formation, although BMP-2 was also stained in occasional PAs without chondroid formation. It is speculated that BMP-2 might be secreted by tumor cells and play a role in chondroid formation in PA by inducing some tumor cells to produce type II collagen and other chondroid matrical substances, like glycosaminoglycans. The expression of BMP-2 is specific to PA and may possibly be used as a useful marker in differentiating PA from other salivary gland tumors.

Topics: Adenocarcinoma; Adenoma; Adenoma, Pleomorphic; Biomarkers, Tumor; Bone Morphogenetic Protein 2; Bone Morphogenetic Proteins; Carcinoma; Collagen; Humans; Immunohistochemistry; Salivary Gland Neoplasms; Transforming Growth Factor beta

It has been found by PCR-SSCP analysis and direct DNA sequencing that a human salivary adenosquamous carcinoma-forming cell line, TYS, has a mutant p53 gene at codon 281Asp-->His. When TYS cells were treated with a differentiation-inducing agent, vesnarinone, cellular proliferation was significantly inhibited on the basis of MTT assay. In addition, it has been found by Northern blotting and/or immunoblotting that expression of p21WAF1 and transforming growth factor-beta (TGF-beta) is up-regulated by treating TYS cells with vesnarinone. TGF-beta 1 alone also induced p21WAF1 expression in TYS cells. Moreover, it has been shown by ELISA that the treatment of TYS cells with vesnarinone results in the enhanced generation of latent TGF-beta 1. The expression of TGF-beta receptor (T beta R), including T beta R-I, T beta R-II and T beta R-III, on TYS cells was detected by affinity cross-linking using 125I-TGF-beta 1 and addition of active TGF-beta 1 into serum-free culture medium inhibited the growth of TYS cells in a concentration-dependent manner. These findings suggest that vesnarinone might directly induce expression of p21WAF1 gene in TYS cells, the product of which may be associated with the inhibition of cell growth and induce differentiation.

Topics: Antineoplastic Agents; Carcinoma, Adenosquamous; Cell Differentiation; Cell Division; Cyclin-Dependent Kinase Inhibitor p21; Cyclins; Gene Expression Regulation, Neoplastic; Genes, p53; Humans; Mutation; Polymerase Chain Reaction; Polymorphism, Single-Stranded Conformational; Pyrazines; Quinolines; Salivary Gland Neoplasms; Transforming Growth Factor beta; Tumor Cells, Cultured