Immunoexpression of HSP27 does not seem to influence the prognosis of oral tongue squamous cell carcinoma.

New methods of early detection and risk assessment have been studied aiming to predict the prognosis of patients and directing a specialized treatment of the oral tongue squamous cell carcinoma (OTSCC). In this context, several molecular biomarkers have been investigated for this purpose, and, among them, the heat shock protein 27 (HSP27) can be named. The study aimed to analyze whether heat shock protein 27 (HSP27) exerts any influence on OTSCC, correlating its immunoexpression with clinicopathological parameters, and patient survival. The sample comprised 55 OTSCC cases and 20 normal oral mucosa specimens. The malignancy grading systems proposed by the WHO in 2005, Brandwein-Gensler et al., and Almangush et al. were applied in a histomorphological study. HSP27 expressions were evaluated through the Immunoreactivity Score System (IRS). Significant values were considered at p <0.05 for all statistical tests. Higher IRS results were observed for normal oral mucosa specimens when compared to OTSCC cases (p <0.001). No significant associations between HSP27 immunostaining, the analyzed clinicopathological parameters and patient survival were observed. The results of the present study indicate lower HSP27 expression in OTSCC cases compared to normal oral mucosa specimens. Thus, HSP27 expression does not seem to influence patient prognosis.

Immunoexpression of HSP27 does not seem to influence the prognosis of oral tongue squamous cell carcinoma

Abstract New methods of early detection and risk assessment have been studied aiming to predict the prognosis of patients and directing a specialized treatment of the oral tongue squamous cell carcinoma (OTSCC). In this context, several molecular biomarkers have been investigated for this purpose, and, among them, the heat shock protein 27 (HSP27) can be named. The study aimed to analyze whether heat shock protein 27 (HSP27) exerts any influence on OTSCC, correlating its immunoexpression with clinicopathological parameters, and patient survival. The sample comprised 55 OTSCC cases and 20 normal oral mucosa specimens. The malignancy grading systems proposed by the WHO in 2005, Brandwein-Gensler et al., and Almangush et al. were applied in a histomorphological study. HSP27 expressions were evaluated through the Immunoreactivity Score System (IRS). Significant values were considered at p <0.05 for all statistical tests. Higher IRS results were observed for normal oral mucosa specimens when compared to OTSCC cases (p <0.001). No significant associations between HSP27 immunostaining, the analyzed clinicopathological parameters and patient survival were observed. The results of the present study indicate lower HSP27 expression in OTSCC cases compared to normal oral mucosa specimens. Thus, HSP27 expression does not seem to influence patient prognosis.

Resumo Novos métodos de detecção precoce e avaliação de risco estão sendo estudados com o intuito de predizer o prognóstico dos pacientes e direcionar um tratamento diferenciado. Neste contexto, vários biomarcadores moleculares têm sido investigados com esta finalidade, dentre eles a heat shock protein 27 (HSP27). Esta pesquisa objetivou analisar se a HSP27 exerce alguma influência nos carcinomas de células escamosas de língua oral (CCELO), correlacionando a sua imunoexpressão com parâmetros clinicopatológicos e com a sobrevida dos pacientes. A amostra foi constituída por 55 casos de CCELO e 20 espécimes de mucosa oral normal. Os sistemas de gradação de malignidade propostos pela OMS em 2005, Brandwein-Gensler et al. e Almangush et al. foram aplicados em um estudo histomorfológico. A expressão da HSP27 foi avaliada através do Sistema de Escore de Imunorreatividade (IRS). Para todos os testes estatísticos foram considerados valores significativos com p<0,05. Foi observado um maior IRS para a mucosa oral normal quando comparado aos casos de CCELO (p<0,001). Não foram encontradas associações significativas entre a imunomarcação da HSP27 com os parâmetros clinicopatológicos analisados e com a sobrevida dos pacientes. Os resultados do presente estudo indicam uma menor expressão da HSP27 nos casos de CCELO quando comparados aos espécimes de mucosa oral normal. Assim, a expressão da HSP27 parece não influenciar o prognóstico dos pacientes.

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.

Análise dos efeitos biológicos associados ao fator de choque térmico 1 (HSF1) no carcinoma de células escamosas oral / Analysis of the biological effects associated with heat shock factor 1 (HSF1) in oral squamous cell carcinoma

O carcinoma de células escamosas oral exibe altas taxas de morbimortalidade e evidências em vários tipos tumorais mostram que processos associados à iniciação, à progressão e à resistência terapêutica são regulados por HSF1. Portanto, esclarecer as vias de participação de HSF1 no câncer oral pode auxiliar no entendimento do seu comportamento biológico. Em uma pesquisa previamente desenvolvida por nosso grupo, foram realizados a análise clinicopatológica e o estudo da imunoexpressão de HSF1 em 70 casos de carcinoma de células escamosas de língua oral (CCELO) em comparação com 30 espécimes de mucosa oral normal (MON). Nesta atual investigação, avaliou-se a participação de HSF1 na tumorigênese do CCELO, através de experimentos in vitro com a linhagem celular SCC15, silenciada e não silenciada, com silenciamento confirmado por qRT-PCR e Western Blot. Foram analisadas a viabilidade e proliferação celular, (CellTiter e BRDU, respectivamente), influência no ciclo celular (iodeto de propideo e análise por citometria de fluxo), capacidade de invasão (sistema transwell/Matrigel)e transição epitélio-mesenquimal (TEM) (expressão de E-caderina e vimentina por qRT-PCR). Nossos resultados anteriores evidenciaram que quanto aos casos de CCELO, 57,1% exibiram estadiamento clínico III ou IV, 82,9% foram gradados como de alto grau segundo Bryne (1998), 47,1% como de alto risco segundo Brandwein-Gensler et al. (2005) e 58,8% como de alto risco de acordo com o modelo BD. Observou-se repercussão da gradação de Bryne (1998) (p= 0,05) na sobrevida livre de doença. Tamanho do tumor T3 ou T4 (p= 0,04), recidiva local (p= 0,02) e modelo BD (p=0,02) repercutiram na sobrevida global. Encontrou-se previamente resultado significativo (p<0,01) quando se comparou a imunoexpressão de HSF1 entre a MON e o CCELO, sem associações significativas da imunoexpressão com os parâmetros clinicopatológicos. A partir dos estudos funcionais, observou-se que HSF1 é superexpresso na linhagem SCC15 comparada aos queratinócitos imortalizados (p<0,005) e que o silenciamento deste gene inibiu a proliferação celular (p< 0,005), avanço nas fases do ciclo celular, com aumento do número de células nas fases G0/G1 (p<0,01) e redução das células na fase S (p<0,001), capacidade de invasão (p<0,05) e TEM, com diminuição da expressão de vimentina (p<0,001) e aumento de E-caderina (p<0,05), quando comparadas as linhagens silenciada e controle. Diante destes resultados, sugere-se que HSF1 pode desempenhar diversas funções que ajudam a manter a estabilidade celular em meio às condições estressoras do microambiente tumoral. Assim, futuramente, estratégias envolvendo sua regulação pode ser uma terapia útil no controle da progressão do câncer oral (AU).

Oral squamous cell carcinoma exhibits high rates of morbimortality and evidence in several tumor types shows that processes associated with initiation, progression and therapeutic resistance are regulated by HSF1. Therefore, to clarify the pathways of HSF1 participation in the oral cancer may help in the understanding of its biological behavior. In research previously developed by our group, a clinicopathological analysis and an immunoexpression study of HSF1 of 70 cases of oral tongue squamous cell carcinoma (OTSCC) were performed in comparison with 30 samples of the normal oral mucosa (NOM). In this current investigation, the role of HSF1 in OTSCC tumorigenesis was evaluated, through in vitro experiments with the SCC15 cell line, silenced and non-silenced, with silencing confirmed by qRT-PCR and Western Blot. Cell viability and proliferation (CellTiter and BrdU, respectively), influence on cell cycle (propidium iodide and flow cytometry analysis), invasion capacity (transwell / Matrigel system), and epithelial-mesenchymal (EMT) (expression of E-cadherin and vimentin by qRT-PCR) were evaluated. Our previous results showed that as for the cases of OTSCC, 57.1% exhibited clinical stage III or IV, 82.9% were graded as high grade according to Bryne (1998), 47.1% as high risk according to BrandweinGensler et al. (2005) and 58.8% as high risk according to the BD model. Bryne's gradation (1998) (p = 0.05) had an impact on disease-free survival. Tumor size T3 or T4 (p = 0.04), local recurrence (p = 0.02) and BD model (p = 0.02) impacted overall survival. A significant initial result (p <0.01) was found when comparing an HSF1 immunoexpression between NOM and OTSCC, with no significant association of immunoexpression with clinicopathological tests. From the functional studies, it was observed that HSF1 is overexpressed in the SCC15 cell line compared to immortalized keratinocytes (p <0.005) and that the silencing of this gene inhibited cell proliferation (p <0.005), advance in the cell cycle phases, with an increase in the number of cells in phases G0/G1 (p <0.01) and reduction of cells in phase S (p <0.001), invasion capacity (p <0.05) and EMT, with decreased vimentin expression (p <0.001) and increased E-cadherin (p <0.05), when compared to silenced and control lines. Given these results, it is suggested that HSF1 can exert a range of functions that maintain cell stability amid the stressful conditions of the tumor microenvironment. Thus, in the future, strategies involving its regulation may be a useful therapeutic tool in controlling the progress of the oral cancer (AU).

Analysis of CD57+ natural killer cells and CD8+ T lymphocytes in periapical granulomas and radicular cysts.

The aim of this study was to compare the number of CD57+ natural killer (NK) cells and CD8+ T lymphocytes between periapical granulomas (PGs) and radicular cysts (RCs). Twenty-fives cases of PGs and 25 of RCs were submitted to histological analysis and immunohistochemistry using anti-CD57 and anti-CD8 biomarkers. Positive cells were counted in 10 fields (400× magnification) and the median value was calculated for each case. Statistical tests were used to evaluate differences in the number of CD57+ NK cells and CD8+ T lymphocytes according to type of lesion, intensity of the infiltrate and thickness of the lining epithelium. The number of CD57+ NK cells and CD8+ T lymphocytes was higher in PGs than in RCs (p = 0.129 and p = 0.541, respectively). Comparison of the number of CD57+ NK cells in atrophic and hyperplastic epithelium revealed a larger number of cells in the atrophic epithelium (p = 0.042). A larger number of CD57+ NK cells and CD8+ T lymphocytes were observed in grade III infiltrates compared to grade I/II (p = 0.145 and p = 0.725, respectively). CD8+ T lymphocytes were more prevalent than CD57+ NK cells in most cases when PGs and RCs were analyzed separately or in combination (p < 0.0001). CD57+ NK cells and CD8+ T lymphocytes play a key role in antiviral defense and the presence of these cells supports evidence suggesting the participation of these microorganisms in the pathogenesis of PGs and RCs. The response mediated by CD8+ T lymphocytes was more frequent, indicating greater participation of the adaptive immunity in these chronic lesions.

Angioleiomioma oral: relato de um caso e revisão dos achados atuais / Oral angioleiomyoma: case report and a review of current findings

Resumo O angioleiomioma é uma neoplasia benigna que, a partir da nova classificação da OMS (2013) para os tumores de tecidos moles, deixou de ser considerado um tumor de origem muscular lisa, passando a ser considerado um tumor de origem perivascular. Raramente os angioleiomiomas ocorrem na cavidade oral. A lesão é tratada cirurgicamente, com prognóstico considerado favorável. Este trabalho revisa os casos de angioleiomioma oral relatados na literatura nos últimos 5 anos e descreve esse tumor em um homem de 44 anos que apresentou um nódulo assintomático localizado em lábio superior, com evolução de 6 meses. As hipóteses diagnósticas foram de adenoma pleomórfico e adenoma canalicular. A lesão foi submetida à biópsia e análise histopatológica e imuno-histoquímica (S100, CD34, α-SMA, H-caldesmon e desmina) confirmaram o diagnóstico de angioleiomioma. Destacamos a imuno-histoquímica como um importante método auxiliar no diagnóstico diferencial do angioleiomioma com outras lesões e, principalmente, com o miopericitoma.

Abstract Angioleiomyoma is a benign neoplasm that was considered a tumor of smooth-muscle origin until the most recent (2013) WHO classification of soft tissue tumors, in which it was reclassified as a tumor of perivascular origin. Angioleiomyomas rarely occur in the oral cavity. These lesions are treated surgically with good prognosis. This article presents a review of reports of oral angioleiomyoma in the literature from the last 5 years and describes the case of a 44-year-old man who presented with an asymptomatic nodule in the upper lip that had developed over a 6-month period. Diagnostic hypotheses of pleomorphic adenoma or canalicular adenoma were raised. Biopsy of the lesion, histopathological and immunohistochemical analysis (S100, CD34, H-caldesmon, and desmin) confirmed a diagnosis of angioleiomyoma. It is noteworthy that immunohistochemistry is an important auxiliary method for differential diagnosis of angioleiomyoma from other tumors, particularly myopericytoma.

Analysis of CD57+ natural killer cells and CD8+ T lymphocytes in periapical granulomas and radicular cysts

Abstract: The aim of this study was to compare the number of CD57+ natural killer (NK) cells and CD8+ T lymphocytes between periapical granulomas (PGs) and radicular cysts (RCs). Twenty-fives cases of PGs and 25 of RCs were submitted to histological analysis and immunohistochemistry using anti-CD57 and anti-CD8 biomarkers. Positive cells were counted in 10 fields (400× magnification) and the median value was calculated for each case. Statistical tests were used to evaluate differences in the number of CD57+ NK cells and CD8+ T lymphocytes according to type of lesion, intensity of the infiltrate and thickness of the lining epithelium. The number of CD57+ NK cells and CD8+ T lymphocytes was higher in PGs than in RCs (p = 0.129 and p = 0.541, respectively). Comparison of the number of CD57+ NK cells in atrophic and hyperplastic epithelium revealed a larger number of cells in the atrophic epithelium (p = 0.042). A larger number of CD57+ NK cells and CD8+ T lymphocytes were observed in grade III infiltrates compared to grade I/II (p = 0.145 and p = 0.725, respectively). CD8+ T lymphocytes were more prevalent than CD57+ NK cells in most cases when PGs and RCs were analyzed separately or in combination (p < 0.0001). CD57+ NK cells and CD8+ T lymphocytes play a key role in antiviral defense and the presence of these cells supports evidence suggesting the participation of these microorganisms in the pathogenesis of PGs and RCs. The response mediated by CD8+ T lymphocytes was more frequent, indicating greater participation of the adaptive immunity in these chronic lesions.

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)