CLIC4 Function in the Epithelial-Mesenchymal Transition of Epithelial Odontogenic Lesions.

BACKGROUND: Odontogenic lesions constitute a heterogeneous group of lesions. CLIC4 protein regulates different cellular processes, including epithelial-mesenchymal transition and fibroblast-myofibroblast transdifferentiation. This study analyzed CLIC4, E-cadherin, Vimentin, and α-SMA immunoexpression in epithelial odontogenic lesions that exhibit different biological behavior. METHODS: It analyzed the immunoexpression of CLIC4, E-cadherin, and Vimentin in the epithelial cells, as well as CLIC4 and α-SMA in the mesenchymal cells, of ameloblastoma (AM) (n = 16), odontogenic keratocyst (OKC) (n = 20), and adenomatoid odontogenic tumor (AOT) (n = 8). Immunoexpressions were categorized as score 0 (0% positive cells), 1 (< 25%), 2 (≥ 25% - < 50%), 3 (≥ 50% - < 75%), or 4 (≥ 75%). RESULTS: Cytoplasmic CLIC4 immunoexpression was higher in AM and AOT (p < 0.001) epithelial cells. Nuclear-cytoplasmic CLIC4 was higher in OKC's epithelial lining (p < 0.001). Membrane (p = 0.012) and membrane-cytoplasmic (p < 0.001) E-cadherin immunoexpression were higher in OKC, while cytoplasmic E-cadherin expression was higher in AM and AOT (p < 0.001). Vimentin immunoexpression was higher in AM and AOT (p < 0.001). Stromal CLIC4 was higher in AM and OKC (p = 0.008). Similarly, α-SMA immunoexpression was higher in AM and OKC (p = 0.037). Correlations in these proteins' immunoexpression were observed in AM and OKC (p < 0.05). CONCLUSIONS: CLIC4 seems to regulate the epithelial-mesenchymal transition, modifying E-cadherin and Vimentin expression. In mesenchymal cells, CLIC4 may play a role in fibroblast-myofibroblast transdifferentiation. CLIC4 may be associated with epithelial odontogenic lesions with aggressive biological behavior.

Does YAP influence cell proliferation and apoptosis in benign epithelial odontogenic lesions?

OBJECTIVE: To analyze the immunohistochemical expression of YAP and its correlation with markers involved in cell proliferation and apoptosis in benign epithelial odontogenic lesions. STUDY DESIGN: The sample consisted of 95 cases of odontogenic lesions (25 dentigerous cysts, 30 non-syndromic odontogenic keratocysts, 30 conventional ameloblastomas, and 10 unicystic ameloblastomas) and 10 dental follicles used as normal odontogenic tissue. The histological sections were submitted to immunohistochemistry with YAP, cyclin D1, Ki-67, and Bcl-2 antibodies. Immunoexpression was analyzed qualitatively and quantitatively using an adapted method. The collected data were analyzed descriptively and statistically (p ≤ 0.05). RESULTS: The highest YAP expression was observed in odontogenic keratocysts, followed by unicystic ameloblastomas and conventional ameloblastomas, which exhibited moderate immunoreactivity predominantly in peripheral cells. Furthermore, significant differences in YAP immunoexpression were observed between the groups analyzed, with significant positive correlations between YAP and cyclin D1 in dentigerous cysts and unicystic ameloblastomas and between YAP and Ki-67 in unicystic ameloblastomas (p < 0.05). However, there were no statistically significant correlations between YAP and Bcl-2 immunoexpression in the groups studied. CONCLUSION: YAP may influence epithelial cell proliferation in odontogenic cysts and tumors, suggesting its possible participation in the progression of the odontogenic lesions studied.

A Systematic Review of Adenoid Ameloblastoma: A Newly Recognized Entity.

BACKGROUND: Recently, a new odontogenic tumor has been described, the so-called adenoid ameloblastoma (AdAM). The aim of this review was to determine the clinical and imaging features of AdAM and to describe its main histopathological findings. METHODS: The systematic review included published cases with a diagnosis of AdAM in the gnathic bones, which had sufficient clinical, imaging, and histopathological data to confirm its diagnosis. The following histopathological diagnostic criteria were adopted: presence of ameloblastoma-like components, duct-like structures, spiral cellular condensations, and a cribriform architecture. RESULTS: Fifteen articles, corresponding to 30 cases of AdAM, were selected. Most cases affected men (63.3%), with a slight preference for the mandible (16:14) and the posterior region of gnathic bones was the most commonly affected site. The mean age at diagnosis was 40.8 years. Clinically, the lesions usually presented as a swelling (53.3%) and, radiographically, as a well-defined radiolucency (33.4%). Surgical resection (40%) was the most frequently adopted treatment and recurrence occurred in 30% of cases. Microscopic examination showed cribriform areas in most AdAM cases (93.3%); duct-like structures and spiral cellular condensations were seen in 100% of the cases. CONCLUSION: The small number of reported cases, the existence of erroneous diagnoses, and the adoption of initial conservative management make it difficult to determine whether AdAM has a higher risk of recurrence or more aggressive biological behavior than conventional ameloblastomas.

Immunohistochemical comparative analysis of tumor stem cell biomarkers in pleomorphic adenoma, adenoid cystic carcinoma and mucoepidermoid carcinoma of salivary glands.

OBJECTIVE: This study aimed to compare the immunoexpression profile of tumor stem cell (TSC) biomarkers CD44, aldehyde dehydrogenase 1 (ALDH1), OCT4, and SOX2 in salivary gland tumors (SGTs). STUDY DESIGN: Sixty tissue specimens of SGTs, including 20 pleomorphic adenomas, 20 adenoid cystic carcinomas (ACCs), and 20 mucoepidermoid carcinomas, in addition to 4 samples of normal glandular tissue, were subjected to immunohistochemistry. The expression of the biomarkers in the parenchyma and stroma was evaluated. Data were analyzed statistically by nonparametric tests (P < .05). RESULTS: Higher parenchymal expression of ALDH1, OCT4, and SOX2 was observed in pleomorphic adenomas, ACCs, and mucoepidermoid carcinomas, respectively. Most ACCs did not express ALDH1. Higher immunoexpression of ALDH1 in major SGTs (P = .021) and of OCT4 in minor SGTs (P = .011) was found. Immunoexpression of SOX2 was related to lesions without myoepithelial differentiation (P < .001) and malignant behavior (P = .002). Furthermore, OCT4 was related to myoepithelial differentiation (P = .009). CD44 expression was related to a better prognosis. Stromal immunoexpressions of CD44, ALDH1, and OCT4 were higher in malignant SGTs. CONCLUSIONS: Our findings suggest the participation of TSCs in the pathogenesis of SGTs. We emphasize the need for further investigations into the presence and role of TSCs in the stroma of these lesions.

Immunohistochemical study of the plasminogen activator system in benign epithelial odontogenic lesions.

The aim of this study was to analyze and compare the immunohistochemical expression of plasminogen activator system (PAS) proteins (uPA, uPAR, and PAI-1) in ameloblastomas (AMBs), odontogenic keratocysts (OKCs), and dental follicles (DFs) representing normal odontogenic tissue, as well as to investigate possible correlations between these proteins. Twenty AMBs, 20 OKCs, and 10 DFs were selected for immunohistochemical analysis. In each case, the immunoexpression of uPA, uPAR, and PAI-1 was evaluated semiquantitatively based on the percentage of positivity in odontogenic epithelial and connective tissue cells. The epithelial immunoexpression of uPA was significantly lower in AMBs when compared to OKCs (p = 0.001) and DFs (p = 0.029). Significantly higher epithelial immunostaining for uPAR was observed in AMBs when compared to OKCs (p < 0.001). There were no significant differences in the epithelial immunoexpression of PAI-1 between AMBs and OKCs (p = 1.000). The correlations found for the expression of the studied proteins were not statistically significant (p > 0.05). However, the epithelial and connective tissue expressions of uPAR have a strong positive and statistically significant correlation in AMBs. The present results suggest that uPA is involved in the pathogenesis of OKCs and that uPAR may participate in tumorigenesis in AMBs. The high percentage of PAI-1-positive cells suggests a possible role for this protein in the development of AMBs and OKCs. Furthermore, the studied proteins do not seem to act synergistically in AMBs, OKCs, and DFs.

Actinic cheilitis in rural workers: prevalence and associated factors.

OBJECTIVE: To evaluate the prevalence of actinic cheilitis in rural workers and factors associated with the development of this condition. METHODS: A cross-sectional study was conducted in a city in Northeastern Brazil. Data were collected by clinical examination and use of a questionnaire validated with 300 rural workers. The χ2 test was employed to identify possible associations between the presence of actinic cheilitis and clinical and demographic variables. Multiple logistic regression analysis was performed using forward stepwise selection. A p value of 0.05 was considered significant. RESULTS: The prevalence of actinic cheilitis was 12.0% in the sample. The highest prevalence of actinic cheilitis was observed in white males, with low educational level, and an approximately 40-year history of sun exposure. Chronic lesions were commonly found in the lower lip and were characterized by scaling, dryness, and mild edema. Skin color, sex, educational level of patients, and cumulative sun exposure (in years), were identified as predictors of development of actinic cheilitis. CONCLUSION: Our results suggest the need to implement educational health strategies aimed to orient the population about risk factors and preventive measures of the disease. Appropriate clinical management of patients with actinic cheilitis is important for prevention of lip cancer.

Role of plasminogen activator inhibitor-1 in oral tongue squamous cell carcinoma: An immunohistochemical and in vitro analysis.

OBJECTIVE: To evaluate the influence of plasminogen activator inhibitor-1 (PAI-1) on the biological behavior and prognosis of oral tongue squamous cell carcinoma (OTSCC). METHODS: Immunoexpression of PAI-1 was analyzed in 60 OTSCC specimens and classified as low-expression (≤50% of positive cells) or high-expression (>50%). In vitro effects of recombinant human PAI-1 (rhPAI-1) were assessed through functional assays on the OTSCC-derived cell line SCC-25. Three cell groups were evaluated: G0 (control), G10 (10 nM rhPAI-1), and G20 (20 nM rhPAI-1). RESULTS: High membrane expression of PAI-1 was associated with tumor budding (p = 0.046) and high-risk cases (p = 0.043). Cytoplasmic and membrane expression of PAI-1 was not associated with patient survival. Cell viability (p = 0.020) and progression to the S-phase of the cell cycle (p = 0.024) were higher in G10 and G20 at 24 h. The percentages of apoptotic/necrotic cells were not affected by rhPAI-1. The presence of rhPAI-1 increased cell migration (p = 0.039) and invasion (p = 0.039) after 24 and 72 h, respectively. CONCLUSION: Our findings indicate the involvement of PAI-1 in the biological behavior of OTSCC, although its expression may not predict patient survival. The in vitro results suggest that PAI-1 stimulates cell proliferation, migration and invasion and may contribute to the aggressive phenotype of OTSCC.

Actinic cheilitis in rural workers: prevalence and associated factors

ABSTRACT Objective To evaluate the prevalence of actinic cheilitis in rural workers and factors associated with the development of this condition. Methods A cross-sectional study was conducted in a city in Northeastern Brazil. Data were collected by clinical examination and use of a questionnaire validated with 300 rural workers. The χ2 test was employed to identify possible associations between the presence of actinic cheilitis and clinical and demographic variables. Multiple logistic regression analysis was performed using forward stepwise selection. A p value of 0.05 was considered significant. Results The prevalence of actinic cheilitis was 12.0% in the sample. The highest prevalence of actinic cheilitis was observed in white males, with low educational level, and an approximately 40-year history of sun exposure. Chronic lesions were commonly found in the lower lip and were characterized by scaling, dryness, and mild edema. Skin color, sex, educational level of patients, and cumulative sun exposure (in years), were identified as predictors of development of actinic cheilitis. Conclusion Our results suggest the need to implement educational health strategies aimed to orient the population about risk factors and preventive measures of the disease. Appropriate clinical management of patients with actinic cheilitis is important for prevention of lip cancer.

Immunohistochemical study of the plasminogen activator system in benign epithelial odontogenic lesions

Abstract: The aim of this study was to analyze and compare the immunohistochemical expression of plasminogen activator system (PAS) proteins (uPA, uPAR, and PAI-1) in ameloblastomas (AMBs), odontogenic keratocysts (OKCs), and dental follicles (DFs) representing normal odontogenic tissue, as well as to investigate possible correlations between these proteins. Twenty AMBs, 20 OKCs, and 10 DFs were selected for immunohistochemical analysis. In each case, the immunoexpression of uPA, uPAR, and PAI-1 was evaluated semiquantitatively based on the percentage of positivity in odontogenic epithelial and connective tissue cells. The epithelial immunoexpression of uPA was significantly lower in AMBs when compared to OKCs (p = 0.001) and DFs (p = 0.029). Significantly higher epithelial immunostaining for uPAR was observed in AMBs when compared to OKCs (p < 0.001). There were no significant differences in the epithelial immunoexpression of PAI-1 between AMBs and OKCs (p = 1.000). The correlations found for the expression of the studied proteins were not statistically significant (p > 0.05). However, the epithelial and connective tissue expressions of uPAR have a strong positive and statistically significant correlation in AMBs. The present results suggest that uPA is involved in the pathogenesis of OKCs and that uPAR may participate in tumorigenesis in AMBs. The high percentage of PAI-1-positive cells suggests a possible role for this protein in the development of AMBs and OKCs. Furthermore, the studied proteins do not seem to act synergistically in AMBs, OKCs, and DFs.

Prognostic Factors and Survival in Adenoid Cystic Carcinoma of the Head and Neck: A Retrospective Clinical and Histopathological Analysis of Patients Seen at a Cancer Center.

Adenoid cystic carcinoma (AdCC) is a rare malignancy that accounts for approximately 1% of all head and neck cancers. This neoplasm is characterized by slow but often relentless growth and dissemination. Our aim was to retrospectively evaluate the clinical-pathological features of patients diagnosed with head and neck AdCC and to identify possible prognostic factors. This retrospective observational study analyzed 87 cases of AdCC of the head and neck. Clinical parameters (tumor size, lymph node and distant metastasis, clinical stage, and survival) were obtained from the records. Survival curves were constructed using the Kaplan-Meier method. A p value ≤ 0.05 was considered significant. There was a slight predominance of cases diagnosed in female patients (54%). The mean age at diagnosis was 51.5 years. Analysis using Cox's proportional hazards model considering 10-year disease-specific survival identified histologic pattern and presence of perineural invasion as independent prognostic variables. Primary tumor size and distant metastasis were prognostic predictors of 5- and 10-year disease-free survival. Detailed analysis of the association between clinical-pathological parameters and prognosis can assist professionals with cancer treatment planning and adequate patient management. Considering the long-term aggressive behavior of AdCC, rigorous patient follow-up is important to identify possible locoregional or distant recurrences.

Recurrence-free survival and prognostic factors of odontogenic keratocyst: a single-center retrospective cohort.

PURPOSE: The aim of the present study was to evaluate the 5-year recurrence-free survival and prognostic factors of odontogenic keratocyst (OKC) from a single-center retrospective cohort in the northeastern region of Brazil. METHODS: Forty cases of OKC comprised the study population. In the cohort analyzed, 18 (45%) cases were recurrent OKCs and 22 (55%) were non-recurrent OKCs. Recurrence-free survival was defined as the period from the release of the histopathological report to the occurrence of relapse or last visit to the service. RESULTS: Comparison of the clinicopathological variables between primary and recurrent OKC lesions revealed no differences in the frequency of epithelial thickness, presence of satellite cysts and cystic spaces, presence of an inflammatory infiltrate, locularity, and lesion borders. The frequency of symptoms was practically the same even after recurrence. Satellite cysts were more frequent in the group of recurrent lesions (n = 9, p = 0.002) and the presence of an inflammatory infiltrate was also significantly associated with recurrent lesions (n = 15, p = 0.006). Previous decompression or marsupialization was associated with recurrence of the lesion (p = 0.010). CONCLUSIONS: In conclusion, the most significant prognostic factors were previous decompression or marsupialization, as well as, morphological parameters associated with the recurrence cases were the presence of an inflammatory infiltrate and satellites cysts. The risk of recurrence is low but continues due to the particularities of epithelial proliferation in OKC.

Role of Twist and Podoplanin in Partial Epithelial-Mesenchymal Transition in Oral Squamous Cell Carcinoma.

The aim of this study was to perform a comparative analysis of podoplanin (PDPN) and Twist immunoexpressions in lower lip and oral tongue squamous cell carcinomas (LLSCC and OTSCC, respectively). PDPN and Twist immunoexpressions were semi-quantitatively evaluated by analyzing the invasion front, the compressive areas, the large islands and nests and dissociated cells of the chosen carcinomas. Their statistical associations and correlations with clinical-pathological characteristics were verified by the Mann-Whitney and Spearman's test. Twist expression was low in both carcinomas, with <25% labeling on the invasive front. Significant differences were observed for LLSCC (p=0.032) and OTSCC (p=0.025) regarding PDPN immunoexpression in relation to the worst invasion patterns determined by a histological malignancy gradation system. Statistically significant negative correlations between PDPN membrane expression and general (r=-0.356, p=0.024) and cytoplasmic Twist expressions (r=-0.336; p=0.034) in LLSCC were also observed. Twist and PDPN are suggested to be associated to a more aggressive invasion pattern in both LLSCC and OTSCC cases but not related to the different biological behaviors on these anatomical sites. Also, it was seen that PDPN membrane expression is inversely related to general and cytoplasmic Twist expression in LLSCC cases.

Caliber Persistent Artery in the Upper Lip: A Case Report with Unusual Histopathological Findings.

Caliber persistent labial artery (CPLA) consists in a dilated portion of the main branch of the labial artery without loss of size. The aim of this study is to report a case of a patient diagnosed with CPLA in the upper lip, emphasizing unusual histopathological and immunohistochemical findings. A 67-year-old female patient with complaint of a pulsating upper lip lesion without painful symptomatology. Under a clinical diagnosis of CPLA, and considering that the patient was edentulous and used a total prosthesis, an excisional biopsy of the lesion was performed to avoid future traumas in the region and consequently possible exuberant local bleeding. At anatomopathological examination structures suggestive of lymphoid follicles and germinal centers were visualized. Immunohistochemistry showed positivity for CD20, CD68, desmin and CD34 and negativity for CD4. The patient did not have a history of allergies, cardiovascular, rheumatic or systemic diseases that could justified the findings. The case presents unusual histopathological structures, evidencing the necessity of more studies about this pathology so scarce in the literature.

Caliber persistent artery in the upper lip: a case report with unusual histopathological findings

Abstract Caliber persistent labial artery (CPLA) consists in a dilated portion of the main branch of the labial artery without loss of size. The aim of this study is to report a case of a patient diagnosed with CPLA in the upper lip, emphasizing unusual histopathological and immunohistochemical findings. A 67-year-old female patient with complaint of a pulsating upper lip lesion without painful symptomatology. Under a clinical diagnosis of CPLA, and considering that the patient was edentulous and used a total prosthesis, an excisional biopsy of the lesion was performed to avoid future traumas in the region and consequently possible exuberant local bleeding. At anatomopathological examination structures suggestive of lymphoid follicles and germinal centers were visualized. Immunohistochemistry showed positivity for CD20, CD68, desmin and CD34 and negativity for CD4. The patient did not have a history of allergies, cardiovascular, rheumatic or systemic diseases that could justified the findings. The case presents unusual histopathological structures, evidencing the necessity of more studies about this pathology so scarce in the literature.

Resumo Artéria labial de calibre persistente (ALCP) consiste em uma parte dilatada do ramo principal da artéria labial que penetra no tecido submucoso sem perda de calibre. O objetivo desse estudo é relatar um caso de uma paciente diagnosticada com ALCP em lábio superior, enfatizando os achados histopatológicos e imuno-histoquímicos incomuns. Paciente de 67 anos, sexo feminino, com queixa de lesão em lábio superior, pulsante, sem sintomatologia dolorosa. Diante do diagnóstico clínico de ALCP, e considerando que a paciente era edêntula e usuária de prótese total, foi realizada biópsia excisional para evitar futuros traumas na região e, consequentemente, sangramento local exuberante. Ao exame anatomopatológico foram visualizadas estruturas sugestivas de folículos linfoides e com formações sugestivas de centros germinativos. No exame imuno-histoquímico observou-se imunopositividade para CD20, CD68, desmina e CD34 e sem imunomarcação para CD4. A paciente relatou não possuir histórico de alergias, doenças cardiovasculares, reumáticas ou sistêmicas que justificassem os achados. O caso apresenta estruturas histopatológicas incomuns, corroborando a necessidade de mais estudos acerca dessa lesão tão pouco discutida na literatura.

Prognostic value of the immunohistochemical detection of epithelial-mesenchymal transition biomarkers in oral epithelial dysplasia: A systematic review

BACKGROUND: Oral potentially malignant disorders (OPMDs) comprise a range of clinical-pathological alterations that are frequently characterized as architectural and cytological derangements upon histological analysis. Epithelial-mesenchymal transition (EMT) has been proposed as a critical mechanism for the acquisition of the malignant phenotype in neoplastic epithelial processes. This study aims to systematically review the current findings on the immunohistochemical expression of epithelial-mesenchymal transition markers in oral potentially malignant disorders and to evaluate their possible application as biomarkers associated with the progression of oral epithelial dysplasias. MATERIAL AND METHODS: A systematic search was performed in the following databases: PubMed, EMBASE, Chinese BioMedical Literature Database, and Cochrane Library. Articles that evaluated the relationship between the expression of EMT markers and the degree of oral epithelial dysplasia were selected for the systematic review. The quality of each eligible study was evaluated by independent reviewers that used operationalized prognostic biomarker reporting guidelines (REMARK). RESULTS: Seventeen articles met all inclusion criteria and were selected. The EMT markers analyzed exhibited an important association with the prognosis of the cases evaluated. The results showed a progressive increase in the expression of nuclear transcription factors and markers of mesenchymal differentiation, as well as negative regulation of epithelial and cell adhesion markers, according to the stage of oral epithelial dysplasia. CONCLUSIONS: The dysregulation of expression of important EMT components in oral dysplastic epithelium is a potential prognostic marker in OPMDs

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Actinic cheilitis: Morphometric parameters and its relationship with the degree of epithelial dysplasia.

Actinic cheilitis (AC) is a potentially malignant lesion caused by chronic sun exposure. This study aimed to evaluate the relationship between the degree of epithelial dysplasia and morphometric findings in AC. Sixty-eight slides of AC cases were selected and classified according to the grade of epithelial dysplasia, following morphologic criteria of World Health Organization. For morphometric analysis, the slides were scanned and images were analyzed using Pannoramic Viewer software. We obtained vertical measurements of the parameters: thicknesses of the keratin layer, lamina propria and zone of solar elastosis in three selected fields. Thirty-seven (54.4%) of the analyzed cases were classified as none/mild dysplasia and 31 (45.6%) as moderate/severe epithelial dysplasia. Cases with a moderate/severe dysplasia exhibited a thicker layer of keratin (median = 0.055 mm) than none/mild dysplasia (median = 0.045 mm) (p = 0.033). No significant differences in the thicknesses of lamina propria and zone of solar elastosis were observed according to the grade of epithelial dysplasia. A positive significant correlation between keratin layer and lamina propria thicknesses was found (p = 0.019). Based on our findings, rigorous clinical follow-up should be recommended for patients whose histopathological examination shows a greater thickness of the keratin layer.

Estudo imuno-histoquímico e in vitro de proteínas do sistema ativador de plasminogênio em carcinomas de células escamosas de língua oral / Immunohistochemical and in vitro study of plasminogen activator system proteins in oral tongue squamous cell carcinomas

O carcinoma de células escamosas (CCE) é a neoplasia maligna mais frequente da cavidade oral e apresenta prognóstico desfavorável. Assim sendo, pesquisas têm buscado esclarecer o papel de biomarcadores no comportamento biológico do CCE oral. Nesta perspectiva, destacam-se o ativador de plasminogênio tipo uroquinase (uPA) e seu receptor (uPAR), além do inibidor do ativador de plasminogênio-1 (PAI-1). O presente trabalho analisou, por meio de imuno-histoquímica, a expressão das proteínas uPA, uPAR e PAI-1 no CCE de língua oral (CCELO) e sua relação com parâmetros clinicopatológicos. Este experimento também avaliou os efeitos in vitro da proteína recombinante humana PAI-1 (rhPAI-1) na linhagem celular SCC25, derivada de CCELO. A imunoexpressão de uPA, uPAR e PAI-1 foi analisada em 60 casos de CCELO, de forma semiquantitativa, nas células neoplásicas do front de invasão tumoral. Visando a associação dos achados imuno-histoquímicos com variáveis clinicopatológicas e taxas de sobrevida, os casos foram classificados nas categorias baixa expressão (≤50% das células positivas) e alta expressão (>50% das células positivas). No experimento in vitro, foram analisados os seguintes grupos: G0 (controle; células cultivadas na ausência de rhPAI-1), G10 (células tratadas com rhPAI-1 a 10 nM) e G20 (células tratadas com rhPAI-1 a 20 nM). Diferenças entre estes grupos foram investigadas através dos ensaios: viabilidade celular (Alamar Blue), ciclo celular (marcação com iodeto de propídio, PI), apoptose/necrose (marcação com Anexina V e PI), atividade migratória (Wound healing) e invasão celular (Transwell). A análise imuno-histoquímica revelou alta expressão do uPA na maioria dos CCELOs, mas sem relações significativas com parâmetros clinicopatológicos. As expressões do uPAR e do PAI-1, em nível membranar, foram associadas a recidivas locais (p=0,019) e ao elevado tumor budding (p=0,046), respectivamente. A expressão membranar do PAI-1 também apresentou associação significativa com o alto escore de risco histopatológico (p=0,043). A análise estatística evidenciou ausência de associações significativas entre as variáveis imunohistoquímicas (uPA, uPAR e PAI-1) e indicadores de prognóstico do CCELO (sobrevida específica e sobrevida livre da doença). No estudo in vitro, decorridas 24 horas da administração da rhPAI-1, os grupos G10 e G20 exibiram maior viabilidade celular em comparação ao grupo controle (p=0,020), assim como aumento da progressão para a fase S do ciclo celular (p=0,024). No que concerne aos percentuais de células apoptóticas e necróticas, não foram encontradas diferenças significativas entre os grupos. Nos grupos celulares cultivados na presença da rhPAI1, também foi constatado aumento da atividade migratória (p=0,039) e do potencial de invasão (p=0,039), respectivamente, nos intervalos de 24 horas e 72 horas. Os achados deste estudo sugerem o envolvimento das proteínas uPA, uPAR e PAI-1 na patogênese do CCELO. Entretanto, a expressão destes biomarcadores pode não estar relacionada com a sobrevida dos pacientes. Os resultados in vitro demonstram que o PAI-1 exerce efeitos estimulatórios na proliferação, migração e invasão celular, podendo assim contribuir para a agressividade biológica do CCELO (AU).

Squamous cell carcinoma (SCC) is the most frequent malignant neoplasm of the oral cavity and has an unfavorable prognosis. Thus, studies have sought to clarify the role of biomarkers in the biological behavior of oral SCC. Within this context, urokinase-type plasminogen activator (uPA) and its receptor (uPAR), as well as plasminogen activator inhibitor 1 (PAI-1), are particularly interesting. The present study analyzed, by means of immunohistochemistry, the expressions of uPA, uPAR and PAI-1 in oral tongue SCC (OTSCC) and their relationship with clinicopathological parameters. This experiment also evaluated the in vitro effects of recombinant human PAI-1 (rhPAI-1) on the OTSCC-derived cell line SCC-25. The immunoexpression of uPA, uPAR and PAI-1 was analyzed semiquantitatively in neoplastic cells of the invasion front of 60 OTSCC cases. Aiming to determine the association between immunohistochemical findings, clinicopathological variables and survival rates, the cases were classified as low expression (≤50% of positive cells) and high expression (>50% of positive cells). The following groups were analyzed in the in vitro experiment: G0 (control; cells cultured in the absence of rhPAI-1), G10 (cells treated with 10 nM rhPAI-1), and G20 (cells treated with 20 nM rhPAI-1). Differences between these groups were investigated using the following assays: cell viability (Alamar Blue), cell cycle (staining with propidium iodide, PI), apoptosis/necrosis (staining with Annexin V and PI), migratory activity (Wound healing), and cell invasion (Transwell). Immunohistochemical analysis revealed high expression of uPA in most OTSCC cases, but there were no significant associations with clinicopathological parameters. The high membrane expression of uPAR and PAI-1 was associated with local recurrence (p=0.019) and high tumor budding (p=0.046), respectively. Membrane expression of PAI-1 also presented a significant association with high-risk cases (p=0,043). Statistical analysis demonstrated no significant associations between the immunohistochemical variables (uPA, uPAR and PAI-1) and prognostic indicators of OTSCC (disease-specific and disease-free survival). In the in vitro experiment, 24 hours after administration of rhPAI-1, G10 and G20 exhibited greater cell viability compared to the control group (p=0.02), as well as increased progression to the S phase of the cell cycle (p=0.024). There were no significant differences in the percentages of apoptotic or necrotic cells between groups. In the groups cultured in the presence of rhPAI-1, migratory activity (p=0.039) and invasion potential (p=0.039) were found to be increased after 24 and 72 hours, respectively. The findings of this study suggest the involvement of uPA, uPAR and PAI-1 in the pathogenesis of OTSCC. Nevertheless, the expression of these biomarkers may not be related to survival of patients. The in vitro results suggest that PAI-1 exerts stimulatory effects on cell proliferation, migration and invasion and may therefore contribute to the biological aggressiveness of OTSCC (AU).

Regulation of Wnt/ß-catenin pathway may be related to Regγ in benign epithelial odontogenic lesions.

OBJECTIVES: The aim of this study was to analyze and compare the immunoexpressions of Regγ, Wnt-1, and ß-catenin in ameloblastomas, adenomatoid odontogenic tumors (AOTs), and odontogenic keratocysts (OKCs). STUDY DESIGN: Thirty solid ameloblastomas, 20 AOTs, and 30 OKCs were selected for analysis of the immunoexpression of Regγ, Wnt-1, and ß-catenin. Each case was semiquantitatively evaluated in the epithelial component and in their different cellular compartments (membrane, cytoplasm, and nucleus). RESULTS: Ameloblastomas displayed higher cytoplasmic and nuclear Regγ expression compared with AOTs and OKCs, as well as higher membrane and cytoplasmic Wnt-1 expression (P < .05). ß-catenin membrane expression was higher in OKCs compared with ameloblastomas and AOTs (P < .05). Nuclear ß-catenin expression was higher in ameloblastomas and AOTs than in OKCs (P < .05). Cytoplasmic and nuclear Regγ expression in AOTs were positively correlated with nuclear ß-catenin expression (P < .05). CONCLUSIONS: The marked expressions of Regγ, Wnt-1, and ß-catenin suggest the participation of these proteins in the pathogenesis of the studied lesions. The greater expressions of Regγ, Wnt-1, and nuclear ß-catenin in ameloblastomas may be related to their more aggressive behavior. Pro-tumor effects of nuclear ß-catenin may be counterbalanced by inhibitory pathways in AOTs, justifying their low aggressiveness.

Immunohistochemical analysis of lymphatic vessel density and mast cells in oral tongue squamous cell carcinoma.

The aim of this study was to analyze lymphangiogenesis and the presence of mast cells in oral tongue squamous cell carcinoma (OTSCC), correlating the findings with clinicopathological parameters (clinical stage, tumor size, nodal metastasis, histological grade of malignancy, local recurrence, and clinical outcome). Fifty-six cases of primary OTSCC were selected. Lymphatic vessels and mast cells were identified by immunostaining with anti-podoplanin (D2-40) and anti-tryptase antibody, respectively. Lymphatic vessel density (LVD) and mast cell density (MCD) were determined in the intratumoral and peritumoral areas. Intratumoral LVD was higher in advanced clinical stages (III/IV) when compared to early-stage (p = 0.017) and in metastatic cases compared to non-metastatic tumors (p = 0.013). Peritumoral LVD and intratumoral or peritumoral MCD did not differ significantly according to the clinicopathological parameters of OTSCCs (p > 0.05). No significant correlations between LVD and MCD were observed at the intratumoral (r = -0.014; p = 0.918) or peritumoral level (r = 0.156; p = 0.251). Our findings suggest that intratumoral lymphatic vessels, compared to peritumoral lymphatic vessels, appear to be more related to the progression of OTSCC. MCD alone does not seem to be determinant for lymphangiogenesis or for the biological behavior of OTSCC, indicating multiple pro- and antitumor effects of these inflammatory cells.

Expression of urokinase-type plasminogen activator and its receptor in squamous cell carcinoma of the oral tongue.

Urokinase-type plasminogen activator (uPA) and its receptor (uPAR) act in the proteolysis of basement membrane and extracellular matrix structures, facilitating tumor invasion. The purpose of this study was to evaluate the relationship between these proteins and clinicopathological parameters in squamous cell carcinoma of the oral tongue (SCCOT). Sixty cases of SCCOT were submitted to immunohistochemistry and analyzed semiquantitatively at the invasion front and in the tumor core. The results were associated with lymph node metastasis, clinical stage, locoregional recurrence, clinical outcome and histological grade of malignancy. A higher expression of uPA was observed in cases of tumors of high-grade versus low-grade malignancy (p = 0.010). Moreover, the cases with the worst pattern of invasion presented an overexpression of uPA (p = 0.011). The presence of locoregional recurrence was associated with uPAR (p = 0.039), and the expression of both biomarkers was much higher at the invasion front than in the tumor core (p < 0.001). The results suggest uPA and uPAR are involved in the progression and aggressiveness of SCCOT, mainly at the tumor-host interface.