Loss of Caveolin-1 Expression in Tumor Cells is Associated with Increased Aggressiveness and Cell Invasion in Oral Squamous Cell Carcinoma.

BACKGROUND: Changes in Caveolin-1 (CAV-1) expression are related to tumorigenesis. The aim of this study was to evaluate the role of CAV-1 in tumor progression in oral squamous cell carcinoma (SCC) tissue samples and the effect of CAV-1 silencing on two oral tongue SCC (OTSCC) cell lines (SCC-25, from a primary tumor, and HSC-3 from lymph node metastases). METHODS: Mycroarray hybridization, mRNA expression, and immunohistochemistry were performed on OSCC tissue samples and corresponding non-tumoral margin tissues. The effects of CAV-1 silencing (siCAV-1) on cell viability, membrane fluidity, on the expression of epithelial to mesenchymal transition (EMT) markers and on cell migration and invasion capacity of OTSCC cell lines were evaluated. RESULTS: Microarray showed a greater CAV-1 expression (1.77-fold) in OSCC tumors than in non-tumoral tissues and 2.0-fold more in less aggressive OSCCs. However, significant differences in CAV-1 gene expression were not seen between tumors and non-tumoral margins nor CAV-1 with any clinicopathological parameters. CAV-1 protein was localized both in carcinoma and in spindle cells of the tumor microenvironment (TME), and CAV-1 positive TME cells were associated with smaller/more aggressive tumors, independent of the carcinoma cells' expression. Silencing of CAV-1 increased cell viability only in SCC-25 cells. It also stimulated the invasion of HSC-3 cells and increased ECAD and BCAT mRNA in these cells; however, the protein levels of the EMT markers were not affected. CONCLUSION: Decreased expression of CAV-1 by tumor cells in OSCC and an increase in the TME were associated with increased cell invasiveness and tumor aggressiveness.

Evaluation of halitosis parameters in patients undergoing head and neck radiotherapy.

OBJECTIVES: To assess halitosis parameters using OralChroma™ and the correlation with salivary flow, oral hygiene index, radiation dose, and tongue-coating index among irradiated head and neck cancer patients compared to patients without cancer. MATERIALS AND METHODS: This cross-sectional study enrolled irradiated and non-irradiated patients divided into two groups. Hydrogen sulfide, methyl mercaptan, and dimethyl sulfide (DMS) levels were measured using a gas chromatograph, and sialometry was performed. The tongue-coating index and simplified oral hygiene index were also assessed. RESULTS: Thirty-eight patients were allocated to each group. Volatile sulfur compound levels were above the thresholds in both groups. Non-irradiated individuals showed higher levels of hydrogen sulfide and dimethyl sulfide. Patients with asialia had an inexpressive tongue-coating index and increased dimethyl sulfide levels. A decrease in salivary flow rate was followed by a significant increase in volatile sulfur compound levels. Higher doses of radiation to the submandibular salivary glands were associated with higher concentrations of sulfide and methyl mercaptan. CONCLUSIONS: Head and neck radiotherapy may be important in the development of halitosis. Irradiated patients with asialia presented insignificant lingual biofilm. Consequently, lower levels of volatile sulfur compounds were detected in this group. Asialia, a severe radiation-induced hyposalivation, impacted the levels of DMS (extraoral origin).

Quantitative analysis of KLF4 and SOX2 expression in oral carcinomas reveals independent association with oral tongue subsite location and histological grade.

BACKGROUND: Stemness factors associated with tumorigenesis in different types of cancers have not been specifically studied in oral tongue SCC (OTSSC). Here, we aimed to quantify expression levels and distribution of KLF4 and SOX2, two relevant stemness factors, in oral SCC including OTSCC samples from different subsites. METHODS AND RESULTS: We determined KLF4 and SOX2 expression levels by immunostaining 35 biopsies of OSCC. Stained wholeslide images were digitized and subjected to automatic cell detection and unbiased quantification using Qupath software. We found statistically significant reduction in KLF4 positive cells density (p= 0.024), and fraction (p= 0.022) in OTSCC from tongue borders compared with other tongue subsites. Instead, quantitative SOX2 analysis did not show differences in expression levels between OTSCC from the borders versus OTSCC developed in others subsites. Notably SOX2 expression was revealed increased in moderately and poorly differentiated OSCC compared with well differentiated ones (positive cells density p= 0.025, fraction p= 0.006). No significant correlation between KLF4 and SOX2 expression was observed, neither in OSCC nor in OTSCC. CONCLUSIONS: KLF4 and SOX2 exhibit opposite expression profiles regarding subsite localization and differentiation level in OSCC. Our study prompts future OTSCC prospective studies looking for clinical prognosis to incorporate detailed subsite information in the analysis.

Stromal categorization in early oral tongue cancer.

Stromal categorization has been used to classify many epithelial cancer types. We assessed the desmoplastic reaction and compared its significance with other stromal characteristics in early (cT1-2N0) oral tongue squamous cell carcinoma (OTSCC). In this multi-institutional study, we included 308 cases treated for early OTSCC at five Finnish university hospitals or at the A.C. Camargo Cancer Center in São Paulo, Brazil. The desmoplastic reaction was classified as immature, intermediate, or mature based on the amount of hyalinized keloid-like collagen and myxoid stroma. We compared the prognostic value of the desmoplastic reaction with a stromal grading system based on tumor-stroma ratio and stromal tumor-infiltrating lymphocytes. We found that a high amount of stroma with a weak infiltration of lymphocytes was associated statistically significantly with a worse disease-free survival with a hazard ratio (HR) of 2.68 (95% CI 1.26-5.69), worse overall survival (HR 2.95, 95% CI 1.69-5.15), and poor disease-specific survival (HR 2.66, 95% CI 1.11-6.33). Tumors having a high amount of stroma with a weak infiltration of lymphocytes were also significantly associated with a high rate of local recurrence (HR 4.13, 95% CI 1.67-10.24), but no significant association was found with lymph node metastasis (HR 1.27, 95% CI 0.37-4.35). Categorization of the stroma based on desmoplastic reaction (immature, intermediate, mature) showed a low prognostic value for early OTSCC in all survival analyses (P > 0.05). In conclusion, categorization of the stroma based on the amount of stroma and its infiltrating lymphocytes shows clinical relevance in early OTSCC superior to categorization based on the maturity of stroma.

Histopathologic grading and its relationship with outcome in oral tongue squamous cell carcinoma.

BACKGROUND: Histopathologic grading has been routinely used as a complement for clinical staging in the prognostication of patients with oral tongue squamous cell carcinoma (OTSCC). However, this subject remains contentious because there is no universally accepted grading system. OBJECTIVES: This study compared the prognostic significance of four histopathologic grading systems in 80 cases of oral tongue squamous cell carcinoma (OTSCC). METHODS: Clinical and follow-up information of the patients were obtained from medical records. Histopathologic malignancy grading of the tumor invasive front, Histologic risk assessment (HRA), World Health Organization (WHO) grading system, and Budding and Depth of invasion (BD) model were evaluated in the surgical specimens. RESULTS: The HRA, histopathologic malignancy grading and WHO systems did not predict survival. Patients with larger tumor size [Hazard ratio (HR): 2.38; 95% confidence interval (CI): 1.07-5.27; P = 0.026] and patients with BD model high-grade tumors (HR: 2.99; 95% CI: 1.03-8.68; P = 0.034) were significantly associated with a poor 5-year overall survival rate. In the multivariate analysis, tumor size was identified as the only significant independent prognostic factor (HR: 2.23; 95% CI: 1.00-4.99; P = 0.050). None of the grading systems studied was associated with 5-year disease-free survival rates. CONCLUSIONS: BD model was the only histopathologic grading system associated with the outcome of patients with OTSCC, indicating its potential value as an effective tool for the prognostication of OTSCC.

Comparison of supervised machine learning classification techniques in prediction of locoregional recurrences in early oral tongue cancer.

BACKGROUND: The proper estimate of the risk of recurrences in early-stage oral tongue squamous cell carcinoma (OTSCC) is mandatory for individual treatment-decision making. However, this remains a challenge even for experienced multidisciplinary centers. OBJECTIVES: We compared the performance of four machine learning (ML) algorithms for predicting the risk of locoregional recurrences in patients with OTSCC. These algorithms were Support Vector Machine (SVM), Naive Bayes (NB), Boosted Decision Tree (BDT), and Decision Forest (DF). MATERIALS AND METHODS: The study cohort comprised 311 cases from the five University Hospitals in Finland and A.C. Camargo Cancer Center, São Paulo, Brazil. For comparison of the algorithms, we used the harmonic mean of precision and recall called F1 score, specificity, and accuracy values. These algorithms and their corresponding permutation feature importance (PFI) with the input parameters were externally tested on 59 new cases. Furthermore, we compared the performance of the algorithm that showed the highest prediction accuracy with the prognostic significance of depth of invasion (DOI). RESULTS: The results showed that the average specificity of all the algorithms was 71% . The SVM showed an accuracy of 68% and F1 score of 0.63, NB an accuracy of 70% and F1 score of 0.64, BDT an accuracy of 81% and F1 score of 0.78, and DF an accuracy of 78% and F1 score of 0.70. Additionally, these algorithms outperformed the DOI-based approach, which gave an accuracy of 63%. With PFI-analysis, there was no significant difference in the overall accuracies of three of the algorithms; PFI-BDT accuracy increased to 83.1%, PFI-DF increased to 80%, PFI-SVM decreased to 64.4%, while PFI-NB accuracy increased significantly to 81.4%. CONCLUSIONS: Our findings show that the best classification accuracy was achieved with the boosted decision tree algorithm. Additionally, these algorithms outperformed the DOI-based approach. Furthermore, with few parameters identified in the PFI analysis, ML technique still showed the ability to predict locoregional recurrence. The application of boosted decision tree machine learning algorithm can stratify OTSCC patients and thus aid in their individual treatment planning.

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).

Análise da expressão imunoistoquímica de regy e de proteínas envolvidas na regulação da apoptose em carcinomas epidermoides de língua oral / Analysis of the immunohistochemical expression of regy and of proteins involved in the regulation of apoptosis in oral tongue squamous cell carcinomas

O câncer na cavidade oral é uma das lesões malignas mais frequentes na população mundial. Como o processo de desenvolvimento das neoplasias malignas remete a danos que promovem um desequilíbrio na regulação da divisão e morte celular, nos últimos anos, diversos estudos foram realizados com o intuito de verificar a influência desses danos no comportamento global das células e na evolução da doença. Nesse contexto, pesquisas recentes mostraram que alterações na expressão de REG podem desempenhar um importante papel na progressão tumoral de várias neoplasias malignas, por interferir na regulação da apoptose. Diante disso, o objetivo desse trabalho foi investigar a expressão imunoistoquímica dos marcadores REG, p53, MDM-2, Bcl-2 e Bax em carcinomas epidermoides de língua (CELs) oral, com a finalidade de promover uma análise comparativa da imunoexpressão destas proteínas com os parâmetros clínico-patológicos de agressividade da lesão, no intuito de identificar se o REG contribui para a progressão do tumor e se interfere na expressão das proteínas relacionadas a apoptose. Para tanto, foram coletadas informações clínicas de 58 pacientes acometidos por CELs. Em seguida, foi realizada análise histopatológica e imunoistoquímica dos marcadores supracitados, em amostras de material biológico parafinado da lesão. Os resultados mostraram que os tumores com metástase nodal e de alto grau histopatológico de malignidade apresentavam percentuais significativamente menores de REG (p<0,05). Em adição, a análise da expressão de p53, MDM-2 e Bax nos diferentes parâmetros clínico-patológicos avaliados nesse trabalho, não revelou diferenças significativas nos percentuais de imunopositividade desses marcadores. Com relação ao Bcl-2, foi visto que tumores de alto grau de malignidade e com óbitos relacionados a doença apresentavam percentual significativamente menor de positividade dessa proteína (p<0,05). Por fim, o teste de correlação de Spearman demonstrou existir fraca correlação positiva, estatisticamente significativa, entre os percentuais de REG e das proteínas MDM-2 e Bcl-2. Baseado nesses achados, pode-se concluir que a redução da expressão de REG parece contribuir para a progressão do CEL oral e pode influenciar na expressão das proteínas relacionadas a regulação da apoptose (AU).

Cancer of the oral cavity is one of the most common malignancies in the world. Since the development of malignant neoplasms is related to damage that causes an imbalance in the regulation of cell division and cell death, many studies have been conducted in recent years to verify the influence of this damage on overall cell behavior and on the progression of the disease. Within this context, recent studies suggest that changes in the expression of REG play an important role in the progression of different malignant tumors by interfering with the regulation of apoptosis. Therefore, the objective of the present study was to investigate the immunohistochemical expression of REG, p53, MDM-2, Bcl-2 and Bax in oral tongue squamous cell carcinoma (OTSCC), correlating the immunoexpression of these proteins with clinicalpathological parameters of tumor aggressiveness, in order to determine whether REG contributes to tumor progression and interferes with the expression of apoptosisrelated proteins. For this purpose, the clinical data of 58 patients with OTSCC were collected and paraffin-embedded tumor specimens were submitted to histopathological analysis and immunohistochemistry using the markers cited above. The results showed significantly higher expression of REG (p<0.05) in low-grade tumors and without lymph node metastases. In addition, comparison of the expression patterns of p53, MDM-2 and Bax according to the clinical-pathological parameters studied revealed no significant differences in the percentage of immunostaining for these markers. Regarding Bcl-2, a significantly lower percentage of immunostaining for this protein was observed in tumors with a high grade of malignancy and tumors associated with deaths related to the disease (p<0.05). Finally, Spearman's correlation test demonstrated a significant weak positive correlation between the percentages of REG and of MDM-2 and Bcl-2. These findings suggest that the reduced expression of REG contributes to the progression of OTSCC and may influence the expression of proteins related to the regulation of apoptosis (AU).

Correlação da imunoexpressão do fator de choque térmico 1 (hsf1) com aspectos clinicopatológicos em carcinomas de células escamosas de língua oral / Immunoexpression correlation of heat shock factor 1 (HSF1) with clinicopathological features in squamous cell carcinomas of tongue

O carcinoma de células escamosas oral apresenta altas taxas de morbidade e mortalidade na população, com isso, enormes esforços estão sendo feitos para categorizar alterações morfológicas e identificar biomarcadores que tenham valor prognóstico, bem como que estratifiquem os pacientes em opções terapêuticas individualizadas. Nessa perspectiva, destaca-se o fator do choque térmico 1 (HSF1), o qual é um fator de transcrição de proteínas do choque térmico (HSPs) que permite ao câncer lidar com estressores associados à malignidade, atuando de diferentes formas na progressão tumoral. Esta pesquisa objetivou realizar a análise clinicopatológica de 70 casos de carcinoma de células escamosas de língua oral (CCELO) e o estudo imunoistoquímico dos níveis de expressão da proteína HSF1 em CCELO em comparação com 30 espécimes de mucosa oral normal (MON), correlacionando-se, ainda, esta imunoexpressão com aspectos clinicopatológicos do CCELO.

Quanto aos casos de CCELO, 57,1% exibiram estadiamento clínico III ou IV, 82,9% foram gradados como de alto grau segundo Bryne (1998) e 47,1% como de alto risco de malignidade segundo Brandwein-Gensler et al., (2005). Foi observada uma taxa de sobrevida livre de doença de 47,84% e taxa de sobrevida global de 68,20% nos casos analisados e que o alto grau de malignidade segundo a Gradação de Bryne (1998) (p= 0,05) e tamanho do tumor T3 ou T4 (p= 0,04), recidiva local (p= 0,02) e invasão perineural (p= 0,02) determinaram impactos negativos nesses tempos de sobrevida. Estes resultados corroboram as informações consolidadas na literatura quanto à influência negativa de alguns indicadores clinicopatológicos na sobrevida dos pacientes com CCELO. Encontrou-se resultado estatisticamente significativo (p<0,01) quando comparou-se a imunoexpressão de HSF1 entre a MON e o CCELO. Esta significativa maior expressão de HSF1 nos casos de CCELO sugere que esta proteína atue, de fato, no processo de patogênese desta lesão. Entretanto, não foram encontradas associações estatisticamente significativas entre esta superexpressão com os parâmetros clínicopatológicos analisados. Esse achado pode refletir a influência de eventos epigenéticos sobre o gene HSF1 ou uma possível estabilidade da expressão desta proteína ao longo da progressão da doença. (AU)

Squamous cell carcinoma of oral tongue shows high rates of morbidity and mortality in the population, therefore, great efforts are being made to classify morphological changes and identify biomarkers that have prognostic value and that are able to group patients in individualized therapeutic options. From this perspective, there is the heat shock factor 1 (HSF1), which is a heat shock factor transcription protein (HSPs) that allows the cancer to deal with stressors associated with malignancy, acting differently in tumor progression. This research aimed to perform a clinico-pathological analysis of 70 cases of oral tongue squamous cell carcinoma (OTSCC) and immunohistochemical study of the expression of HSF1 protein in OTSCC, comparing it with 30 specimens of normal oral mucosa (NOM), and correlating this immunostaining with clinico-pathological aspects of OTSCC. To analyze the association between immunoexpression of HSF1 and clinicophatoloical aspects, the cases were categorized in minor and major overexpression, based in the median immunostaining score.

Regarding the cases of OTSCC, 57.1% showed clinical stage III or IV, 82.9% were graded as high grade according to Bryne (1998) and 47.1% as high risk of malignancy according to Brandwein-Gensler et al., (2005). A disease free survival rate of 47.84% and overall survival rate of 68.20% was observed in the analyzed cases, and the high degree of malignancy according to Bryne’s system (1998) (p=0.05), tumor size T3 or T4 (p=0.04), local recurrence (p=0.02), and perineural invasion (p=0.02) determined negative impacts in survival time. We observed also a statistically significant result (p<0.01) when comparing the immunoreactivity of HSF1 between NOM and OTSCC. This significantly increased expression of HSF1 in cases of OTSCC suggests that this protein acts, indeed, in the pathogenesis of this disease. However, there were no statistically significant associations between this overexpression and the clinico-pathological parameters analyzed. This finding may reflect the influence of epigenetic events on HSF1 gene or a possible stability of this protein expression throughout disease progression. (AU)

Activin A immunoexpression as predictor of occult lymph node metastasis and overall survival in oral tongue squamous cell carcinoma.

BACKGROUND: The presence of regional lymph node metastasis has an important impact on clinical management and prognostication of patients with oral tongue squamous cell carcinoma (SCC). Approximately 30% to 50% of patients with oral tongue SCC have regional metastasis at diagnosis, but the limited sensibility of the current diagnostic methods used for neck staging does not allow detection of all cases, leaving a significant number of undiagnosed metastasis (occult lymph node metastasis). In this study, we evaluated whether clinicopathologic features and immunohistochemical detection of carcinoma-associated fibroblasts (CAFs) and activin A could be predictive markers for occult lymph node metastasis in oral tongue SCC. METHODS: One hundred ten patients with primary oral tongue SCC, who were classified with early stage tumor (stage I and II) and received surgical treatment with elective neck dissection, were enrolled in the study. RESULTS: Among all examined features, only high immunohistochemical expression of activin A was significantly associated with presence of occult lymph node metastasis (p = .006). Multivariate survival analysis using the Cox proportional hazard model showed that the expression of activin A was an independent marker of reduced overall survival with a 5-year survival of 89.7% for patients with low expression compared to 76.5% for those with high expression (hazard ratio [HR], 2.44; 95% confidence interval [CI], 1.55-3.85; p = .012). CONCLUSION: Our results demonstrated that immunodetection of activin A can be useful for prognostication of oral tongue SCC, revealing patients with occult lymph node metastasis and lower overall survival.

A simple novel prognostic model for early stage oral tongue cancer.

The prognostication of patient outcome is one of the greatest challenges in the management of early stage oral tongue squamous cell carcinoma (OTSCC). This study introduces a simple histopathological model for the prognostication of survival in patients with early OTSCC. A total of 311 cases (from Finland and Brazil) with clinically evaluated early stage OTSCC (cT1-T2cN0cM0) were included in this multicentre retrospective study. Tumour budding (B) and depth of invasion (D) were scored on haematoxylin-eosin-stained cancer slides. The cut-off point for tumour budding was set at 5 buds (low <5; high ≥5) and for depth of invasion at 4mm (low <4mm; high ≥4mm). The scores of B and D were combined into one model: the BD predictive model. On multivariate analysis, a high risk score (BD score 2) correlated significantly with loco-regional recurrence (P=0.033) and death due to OTSCC (P<0.001) in early stage OTSCC. The new BD model is a promising prognostic tool to identify those patients with aggressive cases of early stage OTSCC who might benefit from multimodality treatment.