Cholesterol depletion induces mesenchymal properties in oral squamous cell carcinoma cell line.

BACKGROUND: Cholesterol in cell membranes is crucial for cell signaling, adhesion, and migration. Membranes feature cholesterol-rich caveolae with caveolin proteins, playing roles in epithelial-mesenchymal transition and cancer progression. Despite elevated cholesterol levels in tumors, its precise function and the effects of its depletion in oral squamous cell carcinoma remain unclear. The aim of this study was to evaluate the influence of cholesterol depletion in oral squamous cell carcinoma cell line and epithelial-mesenchymal transition process. METHODS: Cholesterol depletion was induced on SCC-9 cells by methyl-ß-cyclodextrin and cell viability, proliferation, apoptosis, and colony formation capacities were evaluated. Gene and protein expressions were evaluated by reverse transcription polymerase chain reaction (RT-qPCR) and Western Blot, respectively, and cell sublocalization was assessed by immunofluorescence. RESULTS: Cholesterol depletion resulted in alteration of oral squamous cell carcinoma cell morphology at different concentrations of methyl-ß-cyclodextrin, as well as decreased cell proliferation and viability rates. Analysis of CAV1 transcript expression revealed increased gene expression in the treated SCC-9 during the 24 h period, at different concentrations of methyl-ß-cyclodextrin: 5 , 7.5, 10, and 15 mM, in relation to parental SCC-9. CAV1 protein expression was increased, with subsequent dose-dependent decrease. A statistically significant difference was observed in samples treated with 5 mM of methyl-ß-cyclodextrin (p = 0.02, Kruskal-Wallis test). The immunofluorescence assay showed lower cytoplasmic and membrane labeling intensity in the treated samples for CAV1. CONCLUSION: These findings indicate the modulation of cholesterol as a possible mechanism underlying the regulation of these molecules and activation of epithelial-mesenchymal transition in oral squamous cell carcinoma.

Potential role of Hedgehog signaling pathway and myofibroblastic differentiation in central giant cell granuloma-A preliminary study.

OBJECTIVE: This study investigated components of the Hedgehog (HH) signaling pathway (SHH, GLI1), cyclin D1, and smooth muscle actin (SMA) in central giant cell granulomas (CGCG). The relationship between these proteins and myofibroblasts was also studied. MATERIAL AND METHODS: Twelve cases of non-aggressive CGCG and 11 cases of aggressive CGCG were studied using immunohistochemistry for SHH, GLI1, Cyclin D1, and SMA. RESULTS: Associations between all proteins in non-aggressive and aggressive CGCG were not significant (P > .05). All cases of CGCG showed significantly higher expression of SMA compared with the other proteins (P < .01). A positive correlation (P = .04) was only observed between SHH and GLI1 for all cases of CGCG. Furthermore, a positive correlation between SHH and GLI1 in non-aggressive CGCG (P = .04) and between GLI1 and cyclin D1 in aggressive CGCG (P = .03) were observed. There was also a negative correlation between the expression of SHH and SMA in non-aggressive CGCG (P = .031). CONCLUSIONS: This study provided insights into the activation of the HH signaling pathway in CGCG. In addition, the activation of this pathway (SHH and GLI1) might play some role in the differentiation of stromal myofibroblasts, although these markers including Cyclin D1 and SMA do not indicate aggressiveness of the CGCG. Furthermore, this myofibroblastic differentiation process would occur at the expense of maturation of these lesions.

The sonic hedgehog signaling pathway contributes to the development of salivary gland neoplasms regardless of perineural infiltration.

The pleomorphic adenoma (PA), mucoepidermoid carcinoma (MEC), and adenoid cystic carcinoma (ACC) are common tumors arising from salivary glands whose histopathology is heterogeneous. The sonic hedgehog signaling pathway (Hh) and signal transducer and activator of transcription 3 (STAT3) play important roles in cell proliferation, favoring tumor growth. The aim of this investigation was to study components of the Hh pathway, as well as STAT3 in salivary gland neoplasms in an attempt to add information about the biological characteristics of these neoplasms. We used 9 cases of PA, 17 cases of ACC, and 20 cases of MEC. Using immunohistochemistry, SHH, GLI1, SUFU, HHIP, and STAT3 were investigated. For comparative purposes, MCM3 (cellular proliferation marker) was also included. In PA, there was high expression of cytoplasmic SHH and SUFU and low expression of STAT3 and MCM3. In the ACC, there was high expression of GLI1, HHIP, and STAT3 and low expression of SHH, SUFU, and MCM3. In the MEC, we observed high expression of SHH, GLI1, SUFU, and HHIP and low expression of STAT3 and MCM3. There was a statistically significant difference between SHH (p = 0.0064), STAT3 (p = 0.0003), and MCM3 (p = 0.0257) when all tumors were compared and a higher expression in parenchyma for all tumors when stroma and parenchyma were compared (p < 0.05). These findings suggests a possible role of Hh pathway in the development and maintenance of the cytoarchitectural pattern of PA, ACC, and MEC, as well as the participation of STAT3 in the development of ACC, irrespective perineural infiltration.

Importance of cone beam computed tomography for diagnosis of calcifying cystic odontogenic tumour associated to odontoma. Report of a case.

The calcifying cystic odontogenic tumour (CCOT) is a rare benign cystic neoplasm not infrequently associated with odontoma. This report documents a case of CCOT associated with compound odontoma arising in the anterior maxilla in a 25-year-old woman. Conventional radiographs showed a large calcified mass with poorly visualized radiolucent margins. The extent and condition of the internal structure of the CCOT associated with odontoma was able to be determined based on radiographic findings from cone beam computed tomography. This advanced image technique proved to be extremely useful in the radiographic assessment of this particular neoplasm of the jawbones.

Simultaneous presentation of juvenile ossifying fibroma in the maxilla and mandible: a case report.

INTRODUCTION: Juvenile ossifying fibroma (JOF) is a controversial and uncommon lesion that has been distinguished from the larger group of ossifying fibromas because of distinct clinical features and some morphological peculiarities. Furthermore, JOF shows an aggressive biological behavior that has led researchers to consider it a benign neoplasm, resulting in its differential diagnosis with important benign and malignant bone neoplasms. PRESENTATION OF CASE: This study describes a case of synchronous presentation of JOF in the mandible and maxilla of a young patient. In addition, the literature was reviewed to identify clinical-pathologic features and possible factors that could help establish the correct diagnosis. A 26-year-old male patient presented simultaneously a lesion affecting the body, angle and ramus of the left mandible and another lesion in the left maxilla. Both lesions were well delimited and radiolucent, being unilocular in the maxilla and multilocular in the mandible. The mandibular lesion was partially resected and the maxillary lesion was submitted to curettage. The diagnosis was JOF. DICUSSION: A delay in seeking medical care and a late diagnosis can have serious consequences for the postoperative functional and esthetic outcome. Much care should be taken during establishment of this diagnosis since an equivocal diagnosis can have serious consequences for the patient in terms of treatment. CONCLUSION: After 1 year, the patient shows no signs or symptoms of recurrence of the lesions and was referred for reconstructive surgery of the mandible. An early and correct diagnosis is necessary to permit the best therapeutic management.

Polypoid non-neural granular cell tumor of the oral cavity: case report and literature review

To report a case of non-neural granular cell tumor (NN-GCT), an uncommon neoplasm, with only six studies worldwide describing cases involving the oral cavity. Methods: A 26-year-old male patient with an erythematous, firm, polypoid nodule in the floor of the mouth that exhibited areas of ulceration and mild bleeding to the touch. A biopsy was performed to aid in the diagnosis. Results: Based on the histopathological and immunohistochemical results (vimentin +, CD68 +, S100 -), the diagnosis was compatible with S100-negative (primitive polypoid non-neural) granular cell tumor. No recurrence was observed over two years of follow-up. Conclusion: The diagnosis of NN-GCT is extremely challenging because this tumor shares histological and immunophenotypic features with many benign and malignant tumors. Although oral NN-GCT may exhibit unusual and atypical histological features, it has an indolent behavior. Thus, until more cases of oral involvement are reported, complete resection and close follow-up are recommended

Prognostic implications of CD44, NANOG, OCT4, and BMI1 expression in tongue squamous cell carcinoma.

BACKGROUND: Tongue squamous cell carcinoma (SCC) contains a cell subpopulation referred to as cancer stem cells (CSCs), which are responsible for tumor growth, metastasis, and resistance to chemotherapy and radiotherapy. The CSC markers have been used to isolate these cells and as biomarkers to predict overall survival. METHODS: The CSC markers CD44, NANOG, OCT4, and BMI1 were investigated using reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) and immunohistochemistry and correlated with clinicopathological parameters. RESULTS: The CD44 overexpression was associated with disease-related death (P = 0.02) and worst prognosis. NANOG was upregulated in nontumoral margins and associated with T1/T2 classification, lymph node metastasis, and worst prognosis. OCT4 was associated with lymph node metastasis and worst overall survival. BMI1 and CD44v3 were overexpressed in tongue SCC. Coexpression of CD44++ /NANOG++ was associated with worst overall survival when compared with patients with CD44-/+ /NANOG-/+ . CONCLUSION: The CSC markers might play an important role not only in CSC trait acquisition but also in tongue SCC development and progression.

Estudo da Expressão imuno-histoquímica da Wnt-5a em displasias epiteliais bucais / Immunohistochemical Study of Wnt-5a Expression in Oral Epithelial Dysplasias

Alterações na expressão das proteínas Wnt no carcinoma espinocelular bucal, permitiram a construção de hipóteses sobre o papel da Wnt-5a em desordens potencialmente malignas. Objetivo: Avaliar a expressão imuno-histoquímica da proteína Wnt-5a em displasias epiteliais bucais. Metodologia: Foram analisados 18 casos de displasia epitelial bucal e 05 casos de tecido bucal sadio oriundos do Serviço de Patologia Bucal da Faculdade de Odontologia da UFBA. A expressão da Wnt-5a foi avaliada de acordo com a localização celular (membranoso, citoplasmático e/ou nuclear) e o índice de marcação estabelecendo-se a intensidade de expressão (ausente, fraca, moderada e forte). Resultados: Não houve maior prevalência entre os gêneros, sendo que a maioria dos pacientes encontrava-se na faixa etária entre 20 e 49 anos (50%). Não houve casos de displasia severa, sendo a maior parte de displasia leve (72,2%). A expressão da proteína Wnt-5a aconteceu principalmente como citoplasmática/nuclear, com a intensidade forte sendo a mais prevalente. Conclusão: O aumento da imuno-expressão da proteína Wnt-5a, observadas nas displasias epiteliais bucais, difere significativamente do padrão da mucosa bucal normal e denota a mudança fenotípica das células epiteliais como parte dos eventos precursores da carcinogênese bucal. Não foi possível verificar a relação entre graduação histológica da displasia epitelial bucal e expressão da Wnt-5a... (AU)

Introduction: Changes in the expression of Wnt proteins in oral squamous cell carcinoma allowed the construction of hypothesis about the role of Wnt-5a in potential malignant disorders. Objective: To evaluate the immunohistochemistry expression of the protein wnt-5a in oral epithelial dysplasia. Method: It was analyzed 18 cases of oral epithelial dysplasia from the Department of Oral Pathology, School of Dentistry (UFBA). The expression of Wnt-5a protein was evaluated as membranous, cytoplasmatic and/or nuclear. The labeling index established the intensity of expression (absent, low, moderate and strong). Results: There was no gender prevalence, being the majority of patients aged between 20 and 49 years (50%). There were no cases of severe dysplasia and mostly was mild dysplasia (72.2%). The cytoplasmic / nuclear expression of Wnt-5a was the most frequent, with the strong intensity being the most prevalent. Conclusion: The increased immuno-expression of the protein Wnt-5a observed in oral epithelial dysplasias, differ significantly from the normal pattern of healthy oral mucosa and shows a phenotypic change of epithelial cells as part of the events precursors of oral carcinogenesis. It was not possible to verify the relation between histological grading of oral epithelial dysplasia and expression of Wnt-5a... (AU)

Elastin accumulation in actinic cheilitis with different degrees of epithelial dysplasia / Acumulación de elastina en queilitis actínica con diferentes grados de displasia epitelial

The extracellular matrix (ECM) plays an important role in the regulation of biological events such as the development of cell migration, proliferation and differentiation. Chronic sun exposure causes changes present in the ECM of actinic cheilitis (AC), a premalignant lesion of the lower lip which helps to understand the carcinogenesis of the lip. This study aimed to investigate elastin, the main component of solar elastosis alternating current in an attempt to establish the relationship between this protein and ECM in epithelial dysplasia. Paraffin embedded tissue sections of the lesions of 35 cases of AC were analyzed by immunohistochemistry for elastin, and became the association with the degree of epithelial dysplasia and age. Highest scores of elastin (+3) was predominant in 45.7 percent of cases of AC, especially in cases of severe dysplasia (n = 3). When comparing the scores of elastin between the different grades of epithelial dysplasia showed no significant difference (P> 0.05, Kruskal-Wallis). This study was not able to demonstrate the influence of elastin on the severity of epithelial dysplasia in AC. Additional studies on other ECM proteins must be conducted in an attempt to better understand the mechanism of malignant progression of the AC.

La matriz extracelular (MEC) juega un papel importante en la regulación de los eventos biológicos, tales como, el desarrollo de la migración celular, proliferación y diferenciación. La exposición solar crónica provoca cambios presentes en la MRC de la queilitis actínica (QA), una lesión premaligna del labio inferior que contribuye a entender la carcinogénesis del labio. Este estudio tuvo como objetivo investigar la elastina, el componente principal de la elastosis solar en corriente alterna en un intento de establecer la relación entre esta proteína y la MEC en displasia epitelial. Se incluyeron en parafina cortes de tejido de las lesiones de 35 casos de QC fueron analizadas mediante técnicas de inmunohistoquímica para elastina, y se hizo la asociación con los grados de displasia epitelial y la edad. La más alta puntuación de la elastina (+3) fue predominante en el 45,7 por ciento de los casos de QA, especialmente en los casos de displasia severa (n = 3). Al comparar las puntuaciones de elastina entre los diferentes grados de displasia epitelial, no mostró diferencia significativa (P> 0,05, Kruskall-Wallis). Este estudio no fue capaz de demostrar la influencia de la elastina sobre gravedad de la displasia epitelial en QA. Estudios adicionales sobre otras proteínas de la MEC deben llevarse a cabo en un intento por comprender mejor el mecanismo de progresión maligna de la QC.

Expressão de transcritos de genes localizados no cromossomo 11q em carcinoma epidermóide de boca e sua relação com critérios de agressividade / Expression of transcripts of genes located on chromosome 11q in squamous cell carcinoma and its relation to criteria of aggressiveness

A instabilidade genética é um importante evento associado ao carcinoma epidermóide de boca, sendo alterações na região cromossômica 11q constantemente relatadas. Neste estudo, genes localizados na região cromossômica 11q, especificamente os genes CTTN, PPFIA1, SHANK2, TAOS1 e MMP-7, foram investigados quanto a diferenças de expressão de transcritos entre carcinomas epidermóides de boca e suas margens correspondentes. A expressão desses genes foi relacionada com aspectos clínicos e histológicos, com critérios de agressividade estabelecidos, e com a sobrevida dos pacientes. Foram analisadas pela técnica de qRT-PCR 29 amostras congeladas de tumores e 25 margens. Todos os genes apresentaram maiores valores de expressão nos tumores em comparação com as margens, embora apenas o gene MMP-7 tenha exibido valores estatisticamente significantes. A expressão do gene MMP-7 mostrou fraca associação com tumores menos agressivos, e os outros genes apresentaram maiores valores de expressão em tumores mais agressivos, sem significância estatística. Não houve diferença estatística entre a freqüência das variáveis clínicas e histopatológicas com a expressão dos genes estudados, porém o PPFIA1 demonstrou maiores níveis de expressão em tumores de assoalho. Em relação à sobrevida, a expressão elevada de PPFIA1 pode implicar em um maior risco de óbito. Assim, é possível a participação do gene MMP-7 no desenvolvimento da neoplasia, e a relação do PPFIA1 com o risco de óbito, porém a expressão de transcritos dos genes CTTN, SHANK2, TAOS1 e MMP-7 não pode ser relacionada com agressividade tumoral e prognóstico.

Genetic instability is an important event associated with oral squamous cell carcinoma, and alterations in the chromosome region 11q are constantly reported. In this study, genes located on chromosome region 11q, specifically genes CTTN, PPFIA1, SHANK2, TAOS1 and MMP-7, were investigated for differences in the expression of transcripts in oral squamous cell carcinoma and their corresponding margins. The expression of these genes was correlated with clinical and histological aspects, aggressiveness criteria established, and with patient survival. Twenty-nine frozen samples of tumors and 25 samples of margin tissue were analyzed using qRT-PCR. All genes showed a higher expression in tumors, compared with the margins, although only the MMP-7 gene demonstrated statistically significant values. The expression of the MMP-7 gene showed weak association with less aggressive tumors, and the other genes showed higher expression in more aggressive tumors, without statistical significance. There was no statistical difference between the frequency of clinical and histopathological variables and the expression of genes studied, however the PPFIA1 gene demonstrated higher levels of expression in tumors of the floor of mouth. With regard to survival, the high expression of PPFIA1 may imply a greater risk of death. Thus, it is possible that the MMP-7 gene participates in the development of malignancy, and PPFIA1 expression may also be associated with risk of death, however, the expression of transcripts of the CTTN, SHANK2, TAOS1 and MMP-7 genes may not be related to tumor aggressiveness and prognosis.