transforming-growth-factor-beta and Tongue-Neoplasms

The aim of this study was to clarify the correlation between imaging findings obtained using intraoral ultrasonography (US) and pathological findings of tongue cancers, and to examine the predictive value of intraoral US findings with respect to occult nodal metastasis. This was a retrospective study based on the medical records of 123 patients with T1-2N0 tongue cancer. The depth of invasion (DOI) on intraoral US was positively correlated with the pathological invasion depth (PID) (ρ = 0.7080, P < 0.0001). Receiver operating characteristic analyses revealed an optimal DOI cut-off value of 4.1 mm and optimal PID cut-off value of 3.9 mm to detect nodal metastasis. Regarding the margin shape of the primary tumour on intraoral US, the incidence of nodal metastasis was significantly higher for the permeated type than for the pressure type (P < 0.001) and wedge-shaped type (P = 0.002). Furthermore, tumours with peritumoural vascularity assessed by power Doppler US had a significantly higher incidence of nodal metastasis than tumours without (P = 0.003). The sensitivity, specificity, and accuracy of the permeated type to predict nodal metastasis was 53.6%, 95.8%, and 86.2%, respectively. These results suggest that intraoral US findings closely reflect pathological findings and could be useful to predict occult nodal metastasis in patients with early-stage tongue cancer.

Topics: Angiography; Humans; Retrospective Studies; Tongue; Tongue Neoplasms; Transforming Growth Factor beta; Ultrasonography

Oral tongue squamous cell carcinoma (OTSCC) is the most frequently encountered malignant epithelial tumors. Semaphorin-7A is a membrane-associated/secreted protein that plays an essential role in the migration and progression of human malignancies. We aimed to investigate the mechanisms of Semaphorin-7A in the growth and migration of OTSCC.. The expressions of Semaphorin-7A in cells were tested by RT-PCR, Western blot, and Immunofluorescence, separately. The activities of OTSCC cells (HSC-3 and Tca8113) were analyzed by MTT, following treatment with Semaphorin-7A or PBS. The migration, invasion, and apoptosis of cells were also determined. The protein expressions of epithelial mesenchymal transition (EMT) pathway were analyzed by Western blot, after treated with Semaphorin-7A in vitro and in vivo. Finally, the mouse model of OTSCC was treated with antibody target for Semaphorin-7A (AntiSema-7A), Semaphorin-7A or PBS, then the tumor size was determined, and histopathological examination and western blot was applied for further confirmation.. In OTSCC cells, Semaphorin-7A was highly expressed, and Semaphorin-7A promoted growth in multiple metastatic OTSCC cell lines. Further study indicated that Semaphorin-7A resulted in up-regulation of Snail, N-cadherin and Vimentin expression, and downregulating of E-cadherin. In addition, The Ets2-repressor factor (ERF) expression was down-regulated, and transforming growth factor (TGF-β)-induced EMT was promoted in OTSCC cells. Then, the proteins of collagen types I (CT-I) and fibronectin (FIB) were also up-regulated after Semaphorin-7A treatment. Furthermore, our results indicated that inhibition of Semaphorin-7A by antibody target for Semaphorin-7A (AntiSema-7A) suppressed OTSCC growth and increased survival in a mouse model of OTSCC. Histopathological examination confirmed the inhibitory effects in vivo.. Semaphorin-7A promoted growth and migration of OTSCC by regulating TGF-β-induced EMT signaling pathway in OTSCC cells, which provided a new interconnection between the Semaphorin-7A and TGF-β-induced EMT signaling pathway.

Topics: Animals; Antigens, CD; Cell Line, Tumor; Cell Movement; Epithelial-Mesenchymal Transition; GPI-Linked Proteins; Humans; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Invasiveness; Random Allocation; Semaphorins; Signal Transduction; Squamous Cell Carcinoma of Head and Neck; Tongue Neoplasms; Transforming Growth Factor beta

The objective of this study was to observe the distribution of regulatory T cells (Tregs) in the development of tongue squamous cell carcinoma (SCC) and to determine the role of Tregs in the progression of tongue SCC. A mouse model of 4-nitroquinoline-1-oxide (4NQO)-induced-tongue SCC was established. The expression of Forkhead box P3 (Foxp3), interleukin 10, transforming growth factor-β, chemokine CC motif ligands 17, 20, and CC chemokine receptor 4 was determined using real-time quantitative polymerase chain reaction. Foxp3 expression was also analyzed using immunohistochemistry. The results were compared with those of control mice and of 4NQO-treated mice treated with a cyclooxygenase-2 (COX-2) inhibitor. Well to moderately differentiated tongue SCC was induced in all of the experimental mice. The amount of Tregs of the experimental mice was over 10 times as much as control mice at the early stage of tumor progression. COX-2 inhibitor did not prevent the progression of tongue SCC and did not reduce the total amount of Tregs. Tregs function at the early stage of the development of tongue SCC, and it may be effective to suppress Tregs at the early stage of tumor progression for the treatment and/or prevention of tongue SCC.

Topics: 4-Nitroquinoline-1-oxide; Animals; Carcinogenesis; Carcinoma, Squamous Cell; Cells, Cultured; Chemokine CCL17; Chemokine CCL20; Disease Models, Animal; Forkhead Transcription Factors; Gene Expression Regulation, Neoplastic; Humans; Interleukin-10; Lymphocyte Count; Male; Mice; Mice, Inbred C57BL; Quinolones; Receptors, CCR4; T-Lymphocytes, Regulatory; Tongue Neoplasms; Transforming Growth Factor beta

Epithelial-to-mesenchymal transition (EMT) is implicated in embryonic development and various pathological events. Transforming growth factor beta (TGFβ) has been reported to induce EMT in tumor cells, which is a critical step in the process of metastasis leading to cancer spreading and treatment failure. However, the involvement of microRNA during the EMT process in tongue squamous cell carcinoma (TSCC) remains to be determined. To address this question, TSCC cell lines SCC9 and CAL27 were treated with human recombinant TGFβ1 for 48 h. miRNA microarray illustrated that miR-639 was significantly downregulated in TGFβ-treated SCC9 cells. Ectopic expression of miR-639 with miRNA mimics effectively blocked TGFβ-induced EMT in SCC9 and CAL27 cells, but inhibition of miR-639 in SCC9 and CAL27 cells with antisense oligonucleotides induced EMT. Computational microRNA target predictions detected a conserved sequence matching to the seed region of miR-639 in the 3'-UTR of FOXC1 mRNA. Luciferase reporter assays revealed that miR-639 targets FOXC1. Ectopic expression of FOXC1 induces EMT in TSCC cells. Silencing FOXC1 expression blocked TGFβ-induced EMT in SCC9 cells. Clinically, reduced miR-639 expression was associated with metastasis in TSCC and poor patient survival. The data from the present study suggest that reduced expression of miR-639 underscores the mechanism of TGFβ-induced EMT in TSCC by targeting FOXC1 and may serve as therapeutic targets in the process of metastasis.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Cell Line, Tumor; Epithelial-Mesenchymal Transition; Female; Forkhead Transcription Factors; Humans; Lymphatic Metastasis; Male; MicroRNAs; Middle Aged; Prognosis; Tongue Neoplasms; Transforming Growth Factor beta

Podoplanin, a transmembrane sialomucin-like glycoprotein, is known to express at high frequency in oral squamous cell carcinomas (OSCC) and possess metastasis-promoting activity such as increased invasion and platelet-aggregating activity. However, the regulatory mechanism of podoplanin expression in OSCC remains unknown.. In the present study, we investigated the podoplanin expression in both clinical specimens from total 80 patients (50 OSCC and 30 pharyngeal SCC) and in 4 OSCC cell lines in vitro.. Immunohistochemical analysis of surgically resected specimens of OSCC revealed podoplanin expression in 70% of OSCC cases with localization primarily in the basal layer of squamous cancer nest and the expression was inversely correlated with squamous cell differentiation. In vitro analysis of OSCC cell lines revealed 36 that podoplanin expression was decreased in response to the squamous cell differentiation (Cytokeratin 10 expression as a marker) induced by treatment with histone deacetylase (HDAC) inhibitors such as sodium butyrate and trichostatin. Furthermore, transforming growth factor-β (TGF-β) significantly enhanced podoplanin expression in OSCC cell lines in line with increased phosphorylation of Smad2. A TGF-β type I receptor inhibitor (SB431542) significantly inhibited such induction of podoplanin expression by TGF-β at both the protein and mRNA level. However, in a subset of OSCC cell line, its expression was only weakly dependent on TGF-β and squamous differentiation.. These results suggest that regulation of podoplanin is not simple, but in the majority of OSCC cell lines, its expression is positively and negatively regulated by TGF-β receptor/Smad signaling pathway and epigenetic mechanism leading to squamous differentiation, respectively.

Topics: Adult; Aged; Aged, 80 and over; Animals; Benzamides; Biomarkers, Tumor; Butyrates; Carcinoma, Squamous Cell; Cell Differentiation; Cell Line, Tumor; Dioxoles; Female; Histone Deacetylase Inhibitors; Humans; Hydroxamic Acids; Immunohistochemistry; Keratin-10; Lymphatic Metastasis; Male; Membrane Glycoproteins; Mice; Mice, Nude; Middle Aged; Mouth Neoplasms; Neoplasm Invasiveness; Pharyngeal Neoplasms; Signal Transduction; Smad2 Protein; Tongue Neoplasms; Transforming Growth Factor beta

This study was performed to determine whether T(H)17 cells are involved in the development and metastasis of head and neck squamous cell carcinomas (HNSCCs).. T(H)17 cells frequencies in 67 HNSCC patients and 21 healthy volunteers were examined by flow cytometric analysis. T(H)17 cell-related cytokines in serum (interleukin (IL) 17, transforming growth factor (TGF) β, and IL-6) were evaluated by using enzyme-linked immunosorbent assay.. It was discovered that the higher frequency of T(H)17 cells was in HNSCCs patients (1.0 ± 0.4%). The cell proportions and related cytokine concentrations were consistent with the tumor TNM stage. The IL-6 concentration showed positive correlation with the frequency of T(H)17 cells (r = 0.661) and IL-17 levels (r = 0.597). The TGF-β concentration showed a positive correlation with IL-17 (r = 0.626) but no relationship with T(H)17 cells (r = 0.431).. The present data suggested that T(H)17 cells may be involved in tumor growth and metastasis of HNSCCs. IL-6 may play an important role in T(H)17 cell differentiation and functions, and TGF-β may be related to IL-17 secretion but not to the differentiation of T(H)17 cells.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Female; Flow Cytometry; Humans; Interleukin-17; Interleukin-6; Lymphatic Metastasis; Lymphocyte Count; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Neoplasm Staging; Th17 Cells; Tongue Neoplasms; Transforming Growth Factor beta

In an orthotopic murine model of head and neck cancer, combined subcutaneous and intratumoral vaccination with recombinant vaccinia virus expressing interleukin-2 (rvv-IL-2) induced significant tumor regression early on therapy. However, its efficacy was restricted by recurrent tumor growth and loco-regional metastases. In this study, we explored the mechanism of tumor metastasis. We compared the levels of expression of a number of molecules involved in tumor metastasis, which included transforming growth factor-beta1 (TGF-beta1), E-cadherin, matrix metalloproteinases (MMPs): MT1-MMP, MMP-2, MMP-9, their tissue inhibitors (TIMPs): TIMP-1/TIMP-2, and pro-angiogenic factors CD31, VEGF-R2, and iNOS between primary and metastatic tumors by real-time RT-PCR and immunohistochemistry. We detected spontaneous lymph node and tongue metastasis. Metastasis was delayed in rvv-IL-2 treated mice. Cultured tumor cells expressed negligible amount of TGF-beta1. Untreated or metastatic tumors, on the other hand, expressed high levels of TGF-beta1 and secreted TGF-beta1 in the sera of tumor-bearing mice. Levels of TGF-beta1 in the sera suddenly jumped at the time when tumor metastasis started. In the metastatic tumors, levels of MT1-MMP, MMP-2, and MMP-9 were significantly elevated (P < 0.001), while levels of TIMP-1/TIMP-2 and E-cadherin were decreased (P < 0.001) compared to control or primary tumors. Levels of CD31, VEGF-R2, and iNOS were also significantly elevated in the metastatic lesions (P < 0.001). The concurrence of high levels of TGF-beta1 in the sera, expression of proteins involved in metastasis and initiation of metastasis suggested possible role of TGF-beta1 in on setting the metastatic cascade in this model.

Topics: Animals; Cadherins; Cell Line, Tumor; Female; Lymph Nodes; Matrix Metalloproteinases; Mice; Mice, Inbred C3H; Mouth Neoplasms; Neoplasm Metastasis; Nitric Oxide Synthase Type II; Platelet Endothelial Cell Adhesion Molecule-1; Reverse Transcriptase Polymerase Chain Reaction; Tissue Inhibitor of Metalloproteinases; Tongue Neoplasms; Transforming Growth Factor beta; Transforming Growth Factor beta1; Vaccination; Vaccinia virus; Vascular Endothelial Growth Factor Receptor-2; Viral Vaccines

Mucoepidermoid carcinoma (MEC) of salivary glands is a malignant, locally aggressive neoplasm with metastatic potential. The clinical course is usually dependent on histology; however, low-grade carcinomas can result in metastases and tumor-related death. Transforming growth factor beta1 (TGF-beta1) is a potent cytokine that affects growth inhibition of various cells and stimulates extracellular matrix production and angiogenesis. Loss of TGF-beta receptor type II (TGF-beta RII) expression has been related to resistance of TGF-beta1-mediated growth control and tumor progression. In this study, we correlate MEC tumor grade with expression of TGF-beta1 and TGF-beta RII.. Immunohistochemical staining was performed on 16 MEC specimens for activated forms of TGF-beta1 and TGF-beta RII. The percentage of cells in which staining yielded positive findings for activated TGF-beta1 and TGF-beta RII was correlated with tumor grade.. Activated TGF-beta1 was detected in 16 specimens (100%) of MEC and showed strong positive and diffuse staining. Predominately cytoplasmic staining of TGF-beta1 was seen in salivary gland ducts, stroma, and endothelial cells. There was an inverse correlation between tumor grade and loss of expression of TGF-beta RII. All low-grade MEC tumors yielded positive staining results, whereas only one case of intermediate-grade MEC had TGF-beta RII expression. No high-grade MEC showed TGF-beta RII expression.. Loss of expression of TGF-beta RII correlates with tumor grade. The localization of activated TGF-beta1 within neoplastic epithelium, tumor-associated stroma, and endothelium suggests that it might play a role in the stromal proliferation and/or angiogenesis associated with MEC.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Carcinoma, Mucoepidermoid; Endothelium; Female; Humans; Immunohistochemistry; Male; Middle Aged; Parotid Neoplasms; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type II; Receptors, Transforming Growth Factor beta; Submandibular Gland Neoplasms; Tongue Neoplasms; Transforming Growth Factor beta; Transforming Growth Factor beta1

Human matrix metalloproteinase-20 (MMP-20, enamelysin) fragments the enamel-specific protein amelogenin and has been shown to be synthesized exclusively by odontoblasts and ameloblasts and in certain odontogenic tumors. Here we demonstrate, for the first time, the expression of MMP-20 mRNA and protein in two carcinoma cell lines originating from the tongue. Treatment of the SCC-25 and HSC-3 cells with phorbol 12-myristate 13-acetate (10 nmol/L) up-regulated MMP-20 mRNA and protein expression by up to 1.6-fold, but transforming growth factor beta (10 ng/mL) had no effect. The latent proform of recombinant (r) human MMP-20 was converted by tumor-related trypsin-2. Activated rMMP-20 did not degrade type I or type II collagen, but efficiently hydrolyzed fibronectin, type IV collagen, laminin-1 and -5, tenascin-C, and beta-casein. This implies that MMP-20 not only participates in dental matrix remodeling but is also present in tongue carcinoma cells.

Topics: Amelogenin; Carcinogens; Carcinoma, Squamous Cell; Caseins; Cell Adhesion Molecules; Cell Line; Collagen Type I; Collagen Type II; Collagen Type IV; Dental Enamel Proteins; Enzyme Precursors; Fibronectins; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Humans; Kalinin; Laminin; Matrix Metalloproteinase 20; Matrix Metalloproteinase Inhibitors; Matrix Metalloproteinases; Recombinant Proteins; RNA, Messenger; Tenascin; Tetradecanoylphorbol Acetate; Tongue Neoplasms; Transforming Growth Factor beta; Trypsin; Tumor Cells, Cultured; Up-Regulation

To investigate the role of TGF-beta on the growth of Tca83 cells.. The cell culture, MTT detection, immunohistochemistry and in situ hybridization detection methods were used.. After 96 h, compared with the control, most groups with low concentrations of TGF-beta (0.2, 1.0, 5.0, 12.5 micrograms/L) showed growth inhibition, while the group with high concentration of TGF-beta (25.0 micrograms/L) showed no growth inhibition. Positive stain of TGF-beta 1, T beta R I mRNA and TGF-beta 1, T beta R I, T beta R II protein could be detected in Tca83 cells.. Tca83 cell line has the potentiality to secret TGF-beta 1. Autocrine and paracrine of TGF-beta 1 could regulate cancer cell's proliferation and differentiation directly. Since the ligand-receptor conduction in Tca83 cell line is intact, external TGF-beta 1 could combine with the T beta R I and T beta R II receptor and exert its inhibitory effect on Tca83 cell line.

Topics: Activin Receptors, Type I; Carcinoma, Squamous Cell; Cell Differentiation; Cell Division; Humans; Immunohistochemistry; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type I; Receptors, Transforming Growth Factor beta; Tongue Neoplasms; Transforming Growth Factor beta; Tumor Cells, Cultured

Retinoids have shown great promise as chemopreventive against the development of squamous cell carcinomas of the upper aerodigestive tract. However, the exact mechanism by which they block new tumors from arising is unknown. Here, we report that 13-cis- and all-trans-retinoic acid, used at clinically achievable doses of 10(-6) mol/L or less, can directly and specifically affect cell lines cultured from oral squamous cell carcinomas, inducing them to switch from an angiogenic to an anti-angiogenic phenotype. Although retinoic-acid-treated and untreated tumor cells make the same amount of interleukin-8, the major inducer of neovascularization produced by such tumor lines, they vary in production of inhibitory activity. Only the retinoic-acid-treated cells produce a potent angio-inhibitory activity that is able to block in vitro migration of endothelial cells toward tumor cell conditioned media and to halt neovascularization induced by such media in the rat cornea. Anti-angiogenic activity is induced in the tumor cells by low doses of retinoids in the absence of toxicity with a kinetics that suggest that it could be contributing to the effectiveness of the retinoids as chemopreventive agents.

Topics: Animals; Breast Neoplasms; Carcinoma, Squamous Cell; Colonic Neoplasms; Cornea; Endothelium, Vascular; Female; Fibrosarcoma; Humans; Interleukin-8; Keratinocytes; Neovascularization, Pathologic; Neovascularization, Physiologic; Neutralization Tests; Phenotype; Rats; Rats, Inbred F344; Tongue Neoplasms; Transforming Growth Factor beta; Tretinoin; Tumor Cells, Cultured