Caracterización de marcadores de células madre tumorales en cáncer escamocelular de cavidad oral

Introducción: El cáncer escamocelular de cavidad oral es una patología con bajas tasas de sobrevivencia. Cuando no es tratado adecuadamente es un tumor de alta recurrencia y resistente al tratamiento. Nuevas hipótesis plantean que las células tumorales progenitoras por sus propiedades de auto renovación, iniciación tumoral, migración y metástasis pueden ser responsables de la manutención y renovación de este tumor. Sin embargo, aún no existe un consenso sobre la verdadera participación de ellas, debido a que su identificación y caracterización es aún un reto experimental. Objetivo: En este trabajo se busca detectar células con expresión de marcadores de células tumorales Progenitoras en muestras cáncer escamocelular de cavidad oral y relacionarlo con los estadios de diferenciación del tumor. Metodología: En esta investigación se tomaron 32 muestras de pacientes con carcinoma escamocelular de cavidad oral. Se logró detectar in situ, mediante la técnica de inmunofluorescencia, cuatro reconocidos marcadores de células tumorales progenitoras. Resultados: Se identificaron los marcadores OCT4, SSEA4, NANOG y TRA-1-60 en los diferentes estadios de diferenciación tumoral, lo que sugiere la participación de las células progenitoras tumorales en la evolución de esta patología. Conclusiones: El establecimiento y correcta identificación de las células tumorales progenitoras abre nuevas vías terapéuticas para el abordaje de este tumor, en busca de mejorar el pronóstico, tasa de sobrevivencia y calidad de vida del paciente.

Introduction: Squamous cell carcinoma of the oral cavity is a pathology with poor survival rates. When it is not adequately treated, it is a tumor with high recurrence and resistance to treatment. According to new hypotheses, progenitor tumor cells, due to their properties of self-renewal, tumor initiation, migration, and metastasis, could be responsible for the maintenance and renewal of this tumor. However, there is still no consensus on their true participation, subsequent to difficult in their identification and characterization. Materials and methods: In this research, 32 samples provided from patients diagnosis with squamous cell carcinoma of the oral cavity were used. To detect specific markers progenitor tumor cells were used immunofluorescence microscopy. Results: The cells markers OCT4, SSEA4, NANOG and TRA-1-60 were identified in the different stages of the tumor samples, all these findings suggest the role of tumor progenitor cells in the evolution of this pathology. Conclusions: The establishment and correct identification of the progenitor tumor cells provide new therapeutic options for the approach of this tumor seeking to improve the prognosis, survival rate and quality of life of the patient.

Concordancia clínica e histopatológica de lesiones blancas en cavidad bucal

Introducción: Las lesiones blancas en cavidad bucal constituyen un problema de salud que responde a causas diversas y afectan a una cifra considerable de personas a nivel mundial. Las condiciones y resultados del diagnóstico clínico de las lesiones sin un basamento histopatológico solo exhiben deducciones empíricas basadas en la experiencia clínica acumulada, que pueden concordar o no con la real naturaleza de la enfermedad diagnosticada. El estudio de su concordancia permite evidenciar la certeza del diagnóstico clínico. Objetivo: Determinar la concordancia clínica e histopatológica de lesiones blancas presentes en cavidad bucal. Métodos: Se realizó un estudio observacional descriptivo de corte transversal en 193 pacientes con lesiones blancas en cavidad bucal seleccionados mediante muestreo no probabilístico de tipo intencionado, que acudieron al Servicio de Estomatología del Policlínico Arturo Puig Ruiz de Villa del municipio Minas, provincia Camagüey, en el período comprendido de enero de 2020 a enero de 2022. Se realizó biopsia de las lesiones para correlacionar el diagnóstico clínico y el diagnóstico histopatológico. Se evaluó la concordancia de los resultados mediante el índice de Kappa. Resultados: Predominó el grupo de edades de 40-49 años, los pacientes más afectados correspondieron al sexo femenino y el color de piel blanca, la lesión que predominó fue la leucoplasia con displasia, el subsitio anatómico más afectado es la mucosa del carrillo y más de la mitad de los pacientes se relacionaron con el tabaquismo como factor de riesgo. La fuerza de concordancia de la leucoplasia fue muy buena respecto al liquen plano y la candidiasis crónica hiperplásica (buena). El carcinoma espinocelular tuvo un nivel de concordancia moderado. Conclusión: Se promediaron los resultados de concordancia para las lesiones blancas presentes en cavidad bucal en los 193 pacientes de la investigación y de manera general el estudio mostró buena concordancia.

Introduction: White lesions in the oral cavity constitute a health problem that responds to diverse causes and affects a considerable number of people worldwide. The conditions and results of the clinical diagnosis of lesions without a histopathological basis only show empirical deductions based on accumulated clinical experience, which may or may not agree with the real nature of the clinically diagnosed entity. The study of their concordance allows evidencing the certainty of the clinical diagnosis. Objective: To determine the clinical and histopathological concordance of white lesions present in the oral cavity. Methods: A cross-sectional descriptive observational study was carried out in 193 patients with white lesions in the oral cavity selected by non-probabilistic purposive sampling who attended the Department of Dentistry of the Arturo Puig Ruiz de Villa Polyclinic from January 2020 to January 2022. The lesions were biopsied to correlate the clinical diagnosis and histopathological diagnosis. The concordance of the results was evaluated using the Kappa index. Results: The predominant age group was 40-49 years, the most affected patients were female and white skin color, the predominant lesion was leukoplakia with dysplasia, the most affected anatomical subsite was the cheek mucosa and more than half of the patients were related to smoking as a risk factor. The concordance strength of leukoplakia was very good with respect to lichen planus and hyperplastic chronic candidiasis (good). Squamous cell carcinoma had a moderate level of concordance. Conclusion: The concordance results for white lesions present in the oral cavity in the 193 patients of the study were averaged and in general the study showed a good concordance.

Comparative Assessment of E-cadherin's Expression between the Metastatic and Non-metastatic Oral Squamous Cell Carcinoma: An Immunohistochemical Study

ABSTRACT Objective: To identify the clinicopathological correlation of E-cadherin expression in metastatic and non-metastatic oral squamous cell carcinoma (OSCC). Material and Methods: A total of 90 paraffin-embedded tissue sections of OSCC were retrieved from the registry. The total selected samples were 45 cases each from the primary lesions of metastatic and non-metastatic OSCC. One section was subjected to routine Hematoxylin and eosin stain and another to immunohistochemical analysis for E-cadherin expression. Results: A non-significant (p˃0.05) increased expression is seen in the non-metastatic group compared to the metastatic group, with predominantly membrane as the staining site in either group. However, the expression of E-cadherin did not reveal any statistically significant association with independent variables such as age, gender, and adverse habits of the patients (p>0.05). On the other hand, with respect to the histological differentiation of OSCC, a significant association (p<0.001) was observed with the well-differentiated type of metastatic OSCC. Conclusion: E-cadherin was useful to some extent in predicting regional metastasis. However, further studies using a panel of biomarkers with increased sample size may help us understand the process involved in metastasis.

Comparison of Clinical and Pathological Staging in Patients with Head and Neck Cancer After Neck Dissection

Abstract Introduction Clinical and pathological staging plays an important role on the prognosis of head and neck cancer (HNC) patients. Objective The present study aims to compare clinical and pathological T, N and overall staging in patients with HNC, to identify factors associated with these discrepancies, and to analyze and compare survival or disease-free survival in staging disagreements. Methods Retrospective cohort including every patient submitted to neck dissection from January 2010 to December 2020 in the department of Otorhinolaryngology of a tertiary hospital center. Results A total of 79 patients were analyzed; their mean age was 58.52 ± 13.15 years old and 88.9% were male. Assessing overall staging, discrepancies were noted in 53% (36.4% upstaging and 16.6% downstaging) and were significantly associated with clinical overall staging (p = 0.006). Regarding T staging, differences were noted in 45.5% (30.3% upstaging and 15.2% downstaging) and were significantly associated with imaging modality (p = 0.016), clinical T staging (p = 0.049), and histology (p = 0.017). Discrepancies in N staging were noted in 38% (25.3% upstaging and 12.7% downstaging) and were significantly associated with age (p = 0.013), clinical N staging (p < 0.001), and presence of extranodal invasion (p < 0.001). Both in Overall, T, and N staging, the aforementioned disagreements were not associated with either higher mortality or higher disease relapse. Conclusion Overall, T, and N staging disagree in an important number of cases, and the overall stage can disagree in up to 53% of the cases. These disagreements do not seem to influence overall and disease-free survival.

Sensitivity and specificity of salivary pipecolic acid in head and neck squamous cell carcinoma

Aim: The aim of the present preliminary case-control study was to test the sensitivity and specificity of salivary pipecolic acid in predicting head and neck squamous cell carcinoma (HNSCC). Methods: High-performance liquid chromatography was used for the analysis of non-stimulated saliva samples from 40 individuals: 20 in the case group (recently diagnosed with untreated HNSCC) and 20 in the control group (individuals without cancer). Both groups included patients taking daily oral hypoglycemic drugs (comorbidity). The case and control groups were matched at a proportion of 1:1 for sex and comorbidity. Results: Mean salivary levels of pipecolic acid were 169.38 ng/ mL in the case group and 114.66 ng/mL in the control group (p<0.001). Individuals who took oral hypoglycemic drugs had higher levels of pipecolic acid in both the case and control groups (p<0.001). The receiver operating characteristic curve analysis revealed 90% sensitivity and 65% specificity for head and neck cancer, with an area under the curve of 0.838 between the case and control groups. Conclusions: Pipecolic acid had high sensitivity for the diagnosis of HNSCC but low specificity in the sample analyzed. Our findings suggest that salivary pipecolic acid levels are associated with glucose homeostasis. Studies with larger samples are required to evaluate the specificity of this metabolite

Lineage and phylogenetic analysis of HPV-16, -18 in saliva of HNSCC patients

Aim: Head and Neck Squamous Cell Carcinoma (HNSCC) is a global health problem whose incidence varies by geographic region and race according to risk factors. Human papillomavirus (HPV) infection is a significant risk factor for HNSCC. HPV-16 and HPV-18 are two forms of HPV that are carcinogenic. HNSCCs that are HPV positive have a better prognosis rather than HPV negative. The purpose of this research was to characterize HPV-16, -18 variations in the saliva of HNSCC patients by examining the genetic diversity of HPV-16, -18 utilizing the full E6, E7, and L1 genes. Methods:The case-control research included 15 patients with HNSCC and 15 healthy volunteers. Unstimulated entire saliva samples were obtained from the case and control groups by spitting method. Genomic DNA was isolated from all saliva samples. A PCR reaction was used to determine the presence of HPV in saliva. HPV-positive samples were genotyped and data were analyzed. We conducted a variant study on the HPV-16, -18 E6, and E7 genes. Results: Three patients with HNSCC were HPV-positive for two HPV genotypes out of 30 people diagnosed with HPV-DNA. HPV-16 and -18 were the most common genotypes. The HPV-16, -18 E6, and E7 genes were sequenced and compared to the HPV-16, -18 (E6, E7) prototype sequence. In all, HPV-16 lineages A1 and HPV-18 lineages A3 were discovered. Conclusion: Regarding the variation of HPV found in Iranian HNSCC patients, the need for further studies in HPV genotyping was seen. Sequencing HPV genes in HNSCC may help answer questions about HPV genotyping in the Iranian population. HPV genotype analysis aids in the development of vaccinations against HNSCC, halting disease progression and preventing HPV-associated HNSCC

Human papillomavirus and head and neck squamous cell carcinoma in a UK population: Is there an association?

Background: Human papillomavirus (HPV) is an evolving important risk factor for head and neck cancer (HNC) especially for individuals who do not smoke and drink alcohol. The aim of this study was to establish the prevalence of HPV infection and elucidate its association with head and neck squamous cell carcinoma (HNSCC) patients in UK population. Methods: The presence and association of HPV was investigated in HNSCC patients in this retrospective clinical study. Samples were obtained from archived biopsies and resections. HPV screening was performed by the use of polymerase chain reaction (PCR) using the GP5+/GP6+ and the SPF1/2 consensus as primers and by immunohistochemistry (IHC). Samples of viral warts that were IHC positive for HPV and fibroepethelial polyps (FEP) were used as positive and negative controls respectively. Results: The cohort included 124 patients with HNSCC with an age range of 27–97 years (median 60 years) and a male to female ratio of 2:1. Among the 124 HNSCC 43/124 (34.7%) were from the tongue 74/124 (60%) presented with advanced stage III or IV disease 112/124 (90%) had a conventional phenotype 84/124 (68%) were moderately differentiated and 89/124 (72%) had bands or cords at the invasive front. Of the 124 patients with HNSCC 84/124 (68%) demonstrated the presence of HPV 0/124 (0%) was for oral squamous cell carcinomas (OSCC). HPV16 was the associated virus type in all positive samples. However no significant association was observed between HPV positivity and other clinico-pathological variables including age and gender of the patients stage and malignancy differentiation. Conclusion: The results we provide suggest that HPV infection is low in HNSCC in general and absent in OSCC specifically in this UK population during this time period. This implies that HPV infection may not play an important role in HNSCC carcinogenesis compared to other risk factors in UK population. This information can aid in more effective treatment approaches for treating UK cases of HNSCC.

Avaliação in vitro e in silico da atividade do S-(-)-Álcool Perílico sobre linhagens celulares do Carcinoma Epidermóide de Língua / In vitro and in silico evaluation of the activity of S-(-)-Peryllic Alcohol on Tongue Squamous Carcinoma cell lines

O carcinoma epidermóide oral (CEO) é a neoplasia maligna mais frequente da cavidade oral e constitui um problema de saúde pública devido a sua alta taxa de incidência e mortalidade devido em muitos casos ao fracasso terapêutico e a resistência tumoral. Assim sendo, destaca-se a busca por novas moléculas biologicamente ativas, como as encontradas nos produtos de origem natural. Este trabalho tem como objetivo avaliar a atividade antineoplásica do S-(-)-álcool perílico (POH) em culturas de células de CEO de língua e predizer sua afinidade através de modelo computacional sobre proteínas que regulam o ciclo celular. Para isso, foram utilizadas duas linhagens celulares de CEO de língua, HSC-3 e SCC-25. Os seguintes grupos foram analisados: G0 (controle; células cultivadas na ausência de POH), G1 (células tratadas com cisplatina a 40 µM), G2 (células tratadas com POH a 0,5 mM), G3 (células tratadas com POH a 1,0 mM), G4 (células tratadas com POH a 1,5 mM) e G5 (células tratadas com POH a 3,0 mM). Diferenças entre estes grupos foram investigadas através dos seguintes ensaios: viabilidade celular (Alamar Blue e Live/Dead assay) e atividade migratória (Wound healing). Foi também realizada a predição de afinidade entre o POH e as moléculas de controle do ciclo celular utilizando a docagem molecular com emprego do software Molegro Virtual Docker, v. 6.0.1. Os dados foram tratados estatisticamente pelo GraphPad Prism 6.0 (GraphPad Software, EUA), análises paramétricas utilizando teste Anova, pós-teste de Tukey e teste estatístico não-paramétricos de Kruskal-Wallis, seguido pelo teste t de estudent foram adotados para determinação de diferenças entre os grupos experimentais. O índice de significância considerado neste trabalho foi de 5%. Para ambas as técnicas de avaliação da viabilidade celular (Alamar Blue e Live/dead assay) analisadas neste trabalho, o POH foi capaz de reduzir a viabilidade celular de linhagens do CEO de língua de maneira dosedependente e tempo-dependente (p<0,05). As concentrações de 1,5 mM e 3 mM do POH obtiveram resultados melhores ou semelhantes aos encontrados na cisplatina 40 µM, para as duas linhagens, na avaliação da viabilidade celular (p<0,05). Os valores de IC50 do POH foram de 1,5 mM para a célula SCC-25 em todos os intervalos de tempo (24 h, 48 h e 72 h), uma vez que, para a linhagem HSC-3, foram de 3 mM para os tempos de 24 h e 48 h e de 1,5 mM para o intervalo de 72 h. O POH foi capaz de inibir a migração das duas linhagens celulares de CEO de maneira dependente da concentração (p≤0,05), comparados ao grupo controle. A habilidade da molécula POH se ligar a proteínas responsáveis pela ativação do ciclo celular foi avaliada usando docking models. Dentre elas, a proteína GTPase Kras mostrou a melhor energia de ligação (-86.70 kcal/mol), apresentando ligações de hidrogênio com os resíduos THR58 (A) e ASP57 (A) e ligações estéricas com os resíduos TRY32 (A) e ALA18 (A). As evidências deste estudo corroboram a ideia de que o POH possui atividade sobre o CEO, sugerindo que essa molécula possa ser uma forte candidata para o desenvolvimento de medicamentos direcionados ao tratamento desta patologia (AU).

Oral squamous cell carcinoma (OSCC) is the most frequent malignant neoplasm of the oral cavity and constitutes a public health problem due to its high incidence and mortality rate caused in many cases by therapeutic failure and tumor resistance. Therefore, the search for new biologically active molecules stands out, such as those found in products of natural origin. This work aims to evaluate the antineoplastic activity of S-(-)-perillyl alcohol (POH) in cell cultures of tongue CEO and to predict its affinity through a computer model on proteins that regulate the cell cycle. For this purpose, two cell lines of tongue CEO were used, HSC-3 and SCC-25. The following groups were analyzed: G0 (control; cells cultured in the absence of POH), G1 (cells treated with 40 µM cisplatin), G2 (cells treated with 0.5 mM POH), G3 (cells treated with 1 .0 mM), G4 (cells treated with 1.5 mM POH) and G5 (cells treated with 3.0 mM POH). Differences between these groups were investigated through the following assays: cell viability (Alamar Blue and Live/Dead assay) and migratory activity (Wound healing). Affinity prediction between POH and cell cycle control molecules were also performed using molecular docking using Molegro Virtual Docker, v. 6.0.1. The data was statistically treated by GraphPad Prism 6.0 (GraphPad Software, USA), parametric analysis using Anova test, Tukey post-test and Kruskal-Wallis non-parametric statistical test, followed by t student test were adopted for determination of differences between the experimental groups. The significance index considered in this work was 5%. For both cell viability assessment techniques (Alamar Blue and Live/dead assay) analyzed in this work, POH was able to reduce the cell viability of tongue CEO lines in a dose-dependent and time-dependent manner (p<0 .05). The concentrations of 1.5 mM and 3 mM of POH obtained better or similar results to those found in 40 µM cisplatin, for the two strains, in the evaluation of cell viability (p<0.05). The IC50 values of POH were 1.5 mM for the SCC-25 cell at all time intervals (24 h, 48 h and 72 h), since for the HSC-3 line they were 3 mM for 24 h and 48 h times and 1.5 mM for the 72 h interval. POH was able to inhibit the migration of the two DSC cell lines in a concentration-dependent manner (p≤0.05), compared to the control group. The ability of the POH molecule to bind to proteins responsible for cell cycle activation was evaluated using docking models. Among them, the protein GTPase Kras showed the best binding energy (-86.70 kcal/mol), featuring hydrogen bonds with residues THR58 (A) and ASP57 (A) and steric bonds with residues TRY32 (A) and ALA18 ( THE). The evidence from this study supports the idea that POH has antineoplastic activity on the CEO, suggesting that this molecule may be a strong candidate for the development of drugs aimed at the treatment of this pathology (AU).

The effect of radiotherapy on survival in newly-diagnosed metastatic head and neck squamous cell carcinoma(HNSCC) / 中华放射肿瘤学杂志

Objective:To evaluate the survival outcomes of radiotherapy in patients with newly-diagnosed metastatic head and neck squamous cell carcinoma (HNSCC) based on data from the Surveillance, Epidemiology and End Results (SEER) database.Methods:A total of 1226 patients newly-diagnosed with metastatic HNSCC between 2010 and 2015 were selected from the SEER database. There were 762 patients (62.1%) in the radiotherapy group and 464 patients (37.9%) in the non-radiotherapy group. Kaplan-Meier method was used to calculate the cancer-specific survival (CSS) and overall survival (OS). The effect of radiotherapy on survival was assessed by Cox multivariate regression and Propensity score-matched analyses (PSM). According to the results of multivariate analysis, the patients were further divided into low-, intermediate- and high-risk groups, and the effect of radiotherapy on survival was analyzed in different risk groups.Results:The median CSS and OS time of the whole group was 11.0 months and 10.0 months, respectively. For patients in the radiotherapy group and non-radiotherapy group, the median CSS time was 13.0 months and 6.0 months, and the median OS time was 12.0 months and 6.0 months, respectively. Multivariate analysis showed that age (CSS, P=0.045;OS, P=0.002), primary tumor site (CSS, P=0.021;OS, P<0.001), T stage (CSS, P=0.001;OS, P=0.002), N stage (CSS, P=0.002;OS, P<0.001), number of metastatic organs (CSS, P<0.001;OS, P<0.001), surgery (CSS, P<0.001;OS, P<0.001), radiotherapy (CSS, P<0.001;OS, P<0.001), and chemotherapy (CSS, P<0.001;OS, P<0.001)were the independent prognostic factors. After PSM, patients with and without radiotherapy in the low-,intermediate-,and high-risk groups, the 3-year CSS rates were 62.5% vs 23.5%( P=0.008), 22.4% vs 15.7%( P=0.001)and 10.5% vs 9.6%( P=0.203), respectively; the 3-year OS were 58.0% vs 20.8%( P=0.002), 19.8% vs 12.7%( P=0.001)and 7.0% vs 6.1%( P=0.166), respectively. Conclusion:Radiotherapy significantly improves CSS and OS in the low- and intermediate-risk groups, but patients in the high-risk group do not benefit from radiotherapy.

Research progress of circular RNA in head and neck squamous cell carcinoma / 中国医师杂志

CircRNA(circular RNA) is a new class of covalently closed circular non-coding RNAs, with the function of the microRNA sponge, regulation of gene expression, and other functions. Studies have confirmed that circRNAs are involved in the occurrence and progression of a variety of tumors, and can be used as biomarkers and therapeutic targets for tumor diagnosis and prognosis evaluation. In this paper, the expression and mechanism of circRNA in head and neck squamous cell carcinoma are reviewed.

Immunoexpression of PD-L1, CD4+ and CD8+ cell infiltrates and tumor-infiltrating lymphocytes (TILs) in the microenvironment of actinic cheilitis and lower lip squamous cell carcinoma

Abstract Lower lip squamous cell carcinomas (LLSCC) could be associated with a previous history of potentially malignant oral diseases (PMOD), especially actinic cheilitis (AC), with high sun exposure being a well-described risk factor. Immune evasion mechanisms, such as the PD-1/PD-L1 (programmed cell death protein 1/programmed death-ligand 1) pathway has been gaining prominence since immunotherapy with immune checkpoint inhibitors showed a positive effect on the survival of patients with different types of neoplasms. Concomitant with the characterization of the tumor microenvironment, the expression of either or both PD-1 and PD-L1 molecules may estimate mutual relations of progression or regression of the carcinoma and prognostic values of the patient. Objective: Considering the importance of tumor microenvironment characterization, this study aims to determine the immunoexpression of PD-L1 and correlate with the frequency of CD4+ and CD8+ cells in AC and LLSCC lesions and with tumor-infiltrating lymphocytes (TILs) in LLSCC and its relationship with histopathological characteristics. Methodology: This sample includes 33 cases of AC and 17 cases of LLSCC. The cases were submitted to histopathological analysis and to CD4+, CD8+, and PD-L1+ cell determination by immunohistochemistry. Results: There was a significant difference among the frequencies of CD4+, CD8+, and PD-L1+ cells between AC and LSCC cases, higher in the last group. Moreover, histopathological and atypical changes in AC and LLSCC were correlated with the frequencies of PD-L1+, CD4+, and CD8+ cells. In AC, PD-L1+ cases had a low frequency of CD4+ cells, but on the other hand, PD-L1+ cases of LLSCC had a higher frequency of CD4+ and CD8+ cells. Conclusion: Therefore, the PD-L1 molecule may be a potential escape route for the immune response in oral lesions, but the mechanisms differ between AC and LLSCC. Future studies related to immune evasion and immunotherapy in oral lesions should consider the analysis of inflammatory infiltrate and TILs.

Clear cell variant of oral squamous cell carcinoma: case report and review

Abstract The clear cell variant of Oral Squamous Cell Carcinoma (OSCC) is an uncommon histological variant. Kuo first discovered it in the skin, and Frazier et al. found it in the oral cavity. We know of only nine cases of clear cell variant of OSCC reported in the literature till now. The present case is in a 60-year-old male patient with an ulcer on the left posterolateral border of the tongue. The patient had a history of chewing tobacco for 22 years. Clinical examination showed features of malignant ulcer associated with pain and discomfort. Histopathological examination revealed sheets and islands of atypical epithelial cells with clear cytoplasm, nuclear and cellular pleomorphism, and few keratin pearls in the connective tissue suggesting OSCC. Various special stains were performed to identify clear cells. Periodic Acid Schiff-Diastase (PAS-D) and Mucicarmine stains showed positive and negative reactions in clear cells, respectively. Immunohistochemical (IHC) analysis for cytokeratin (AE1/AE3) showed diffuse positivity in clear cells and other epithelial cells. Based on special stains and IHC markers, we confirmed the diagnosis as a clear cell variant of OSCC. This variant is rare and presents diagnostic challenges. It is said to be aggressive in nature. More such cases should be reported to understand its biological behavior and prognosis.

Secondary primary tumor mimicking osteoradionecrosis

ABSTRACT Nasopharyngeal carcinoma (NPC) is a malignant tumor rarely found in the head and neck, representing about 1% of all malignancies. The main treatment for NPC is radiation therapy, which is often given in combination with chemotherapy. However, such treatment may lead to long‐term complications, including second primary tumors (SPTs) and osteoradionecrosis (ORN). Both complications have similar radiological characteristics, which can lead to erroneous diagnoses. This paper describes a case of a second primary tumor in a patient after 20 years of radiotherapy in the area where a previous extraction was performed, mimicking an osteoradionecrosis process.

X-radiation does not affect the dental pulp: a morphological study in rats / A radiação X não afeta a polpa dentária: um estudo morfológico em ratos

Introduction: Radiotherapy is one of the methods used as a treatment for malignant tumors in the head and neck region and it can cause tissue damage in the irradiated areas. In head and neck radiotherapy, teeth are often included within the irradiation area and, consequently, the dental pulp; which receives high doses of radiation. Objective: To evaluate the effects of ionizing radiation on the pulp tissue of rat teeth. Methodology: A double-blind experimental assay with 35 Albinus Wistar rats divided into seven groups was performed; one control group, three groups irradiated with 15 Gy, and three groups irradiated with 25 Gy. The irradiated groups were submit-ted to a single dose of radiation and sacrificed 24 hours, 7 days, and 22 days after irradiation, respectively. The samples were evaluated for the morphological presence of inflammatory infiltrate, edema, necrosis, fibrosis, and degeneration of blood vessels. Statistical analysis was performed using the Kruskal-Wallis and Dunn tests with p < 0.05. Results: Hyaline degeneration of the pulp blood vessels in the irradiated teeth was statistically signifi-cant in all irradiated groups. Inflammatory infiltrate, edema, necrosis or fibrosis was not observed. Conclusion:A single X-radiation dose is not able to affect the dental pulp connective tissue in the long term with no clinical damage.

Introdução: A radioterapia é um dos métodos utilizados como tratamento para tumores malignos em região de cabeça e pescoço e que pode causar danos aos tecidos nas áreas irradiadas. Na radioterapia de cabeça e pescoço, os dentes são comumente incluídos dentro da área de radiação e, consequentemente, a polpa dentária, recebe altas doses de radiação. Objetivo: Avaliar os efeitos da radiação ionizante no tecido pulpar de dentes de ratos. Metodologia: Foi realizado um ensaio experimental duplo-cego com 35 ratos Albinus Wistar divididos em sete grupos: um grupo controle, três grupos irradiados com 15 Gy e três grupos irradiados com 25 Gy. Os grupos irradiados foram submetidos a uma dose única de radiação e sacrificados 24 horas, 7 dias e 22 dias após a irradiação, respectivamente. As amostras foram avaliadas quanto à presença morfológica de infiltrado inflamatório, edema, necrose, fibrose e degeneração nos vasos sanguíneos. A análise estatística foi realizada por meio dos testes de Kruskal-Wallis e Dunn com p < 0.05. Resultados: Degeneração hialina nos vasos sanguíneos pulpares dos dentes irradiados foi estatisticamente significante em todos os grupos irradiados. Não foi observado infiltrado inflamatório, edema, necrose ou fibrose. Conclusão: Uma dose única de radiação X não é capaz de afetar o tecido conjuntivo da polpa dentária a longo prazo sem danos clínicos.

Tinción con azul de toluidina en biopsia dirigida de lesiones displásicas de la mucosa bucal: informe de casos clínicos / Toluidine blue guided biopsy of dysplastic lesions of the oral mucosa: clinical cases report

Objetivo: Los desórdenes de mucosa bucal potencialmente malignos pueden presentar áreas displásicas. En estos casos, la biopsia es un procedimiento imprescindible para un correcto diagnóstico. La inspección visual y la palpación, como método de selección del área de biopsia, ofrecen sensibilidad y especificidad adecuadas pero mejorables. El objetivo de este artículo es presentar una serie de casos clínicos en los que se describen el empleo y la interpretación de la tinción vital con azul de toluidina como método complementario para contribuir a una mejor elección del área de biopsia. Casos clínicos: Se trata de siete casos de lesiones con sospecha de displasia epitelial en mucosa bucal. En cada uno se detalla la correlación de las áreas teñidas con las manifestaciones clínicas y con el diagnóstico de displasia. Además, se muestran patrones de tinción considerados falsos positivos. En la interpretación de la tinción positiva, se tuvieron en cuenta el aspecto superficial y el color de la lesión teñida. El empleo combinado de inspección, palpación y tinción vital podría constituir un procedimiento integral de utilidad para obtener mayor precisión en la determinación del sitio de biopsia en comparación con los mismos procedimientos aplicados de manera individual. En la interpretación de la tinción positiva con azul de toluidina deberían considerarse el aspecto superficial y el color de la lesión teñida (AU)

Aim: Potentially Malignant Disorders in the oral cavity can present dysplastic areas. In these cases, the biopsy is an essential procedure for a correct diagnosis. Visual inspection and palpation, are adequate methods to select the area for the biopsy, however there is margin for improvement. The objective of this article is to present a series of clinical cases in which the use and interpretation of vital staining with Toluidine Blue is described as a complementary method to contribute to a better choice of the biopsy area. Clinical cases: Seven clinical cases that presented lesions with suspected epithelial dysplasia in the oral mucosa were presented. The correlation of the stained areas with the clinical manifestations and with the diagnosis of dysplasia is detailed in each case. Staining patterns considered false positives are also shown. In the interpretation of the positive staining, the superficial appearance and color of the stained lesion were considered. The combined use of inspection, palpation and vital staining could constitute a useful comprehensive procedure to obtain greater precision in determining the biopsy site in relation to the same procedures applied individually. In the interpretation of the positive staining with Toluidine Blue, the superficial appearance and color of the stained lesion should be considered (AU)

Post-radiotherapy recurrence of conventional oral squamous cell carcinoma showing sarcomatoid components: an immunohistochemical study

Spindle cell squamous cell carcinoma (SpSCC) is a rare biphasic malignant neoplasm, uncommonly affecting the oral cavity. The SpSCC diagnosis is difficult, especially when it exhibits inconspicuous morphology, inadequate tissue sampling, or association with an exuberant inflammatory reaction. Post-radiotherapy recurrent SpSCC occurring at the same site of conventional SCC is a rare phenomenon. A 59-year-old man was complained of "painful injury on the tongue" with 20 days of duration. He reported smoking and alcohol consumption. Medical history revealed conventional SCC on the tongue treated with surgery and radiotherapy 10 years ago. Intraoral examination showed a polypoid lesion with ulcerated areas, measuring 3 cm in diameter, on the tongue and floor of the mouth, at the same site of previous conventional SCC. The microscopical analysis showed small foci of carcinomatous component admixed with an exuberant inflammatory reaction. Immunohistochemistry highlighted the sarcomatoid component. Both malignant components were positive for EMA, CD138, p40 (deltaNp63), p63, and p53. Moreover, CK AE1/AE3 evidenced the carcinomatous component, whereas vimentin stained the sarcomatoid component. The Ki-67 was >10%. The current case emphasizes the importance of immunohistochemistry in the differential diagnosis of SpSCC from mimics and documents a rare complication of Ionizing Radiation.

Comparison of altered fractionation schedule with concurrent chemo-radiation for squamous cell carcinoma of head and neck

Background: Aim of the study was to compare the response of altered fractionation schedule with concurrent chemo-radiation in patients with primary and the nodal disease.Methods: Total of 40 patients (20 in each arm) with stage 1- 4 squamous cell carcinoma of the head and neck with a performance status of 0-2 (ECOG) were included in the study. Arm A was altered fractionation schedule where in patients received 6 fractions per week to a total dose of 6600 cGy in 33 fractions. In Arm B, patients received conventional radiotherapy with concurrent chemotherapy three weekly Inj. of cisplatin (100 mg/m2). Patients were evaluated for acute toxicity every week using the Acute Radiation Morbidity Scoring Criteria. The response was assessed after 6 weeks and 12 weeks post treatment using the RECIST criteria. Data was statistically analyzed.Results: Seventeen patients in Arm A and 18 patients in Arm B completed the treatment. At the end of three months, In Arm A, 7 patients had complete response and in Arm B, 9 patients had complete response of the primary (p>0.05).  When the complete nodal response was compared in both the arms, there was no difference (2 vs 4 in Arm A vs Arm B resp.). But there were more partial nodal responders in Arm B (p = 0.016). The acute toxicities were comparable in both the arms.Conclusions: Altered fraction radiotherapy can be used in early lesions with minimal nodal burden but with locally advanced disease or large nodal burden addition of chemotherapy should not be avoided.

EBV-negative lymphoepithelial-like carcinoma of the lower lip

Lymphoepithelial-like carcinoma (LEC) is a rare malignant neoplasm, which can be associated with Epstein-Barr virus (EBV) infection. Histologically, LEC is an undifferentiated carcinoma with an intermixed reactive lymphoplasmacytic infiltrate. LEC appears to be an uncommon tumor type of lip carcinoma. An 82-year-old white woman presented a lesion on her lower lip that developed over the last year. The lesion was characterized by ulceration with flat edges, hardened base, painful, and absence of regional lymphadenopathy. Microscopical analysis evidenced an intense inflammatory infiltrate, composed of lymphoplasmacytic cells, associated with scarce pleomorphic epithelial cells. Immunohistochemistry highlighted the LEC cells with strong expression of pan-CK AE1/AE3, EMA, p63, and p53. CD138 was also faintly positive. Ki-67 was >85%. In situ hybridization analysis did not show evidence of EBV. A diagnostic of EBV-negative LEC was made. We present an uncommon type of lip carcinoma, which can represent a diagnostic challenge for clinicians and pathologists.

Beta-adrenergic pathway activation in head and neck cancer: a comprehensive analysis of the clinical and genomic features using integrated bioinformatics / Ativação da via beta-adrenérgica no câncer de cabeça e pescoço: uma análise abrangente das características clínicas e gênomicas utilizando a bioinformática

O estresse crônico leva à ativação da via de sinalização beta-adrenérgica. Sua ativação tem sido implicada na progressão de diferentes tipos de câncer, mas seu papel nos carcinomas espinocelulares de cabeça e pescoço (CECPs) permanece indefinido. O objetivo deste estudo foi investigar o papel da ativação da via betaadrenérgica na progressão dos CECPs, avaliar seu impacto na sobrevida dos pacientes e buscar possíveis terapias para pacientes que encontravam-se com a via beta-adrenérgica ativa. Quinhentos e vinte pacientes do The Cancer Genome Atlas com CECPs primários foram divididos em dois grupos: ADRB2baixa / SLC6A2baixa e ADRB2alta / SLC6A2alta. A associação de características clinicopatológicas e genômicas entre os grupos foram analisadas utilizando bioinformática. Os genes diferencialmente expressos (DEGs) foram identificados através da análise da expressão diferencial. A análise de sobrevida também foi realizada com base nas expressões ADRB2 e SLC6A2. Foram identificados medicamentos em potencial para tratamento de CECPs com base nos DEGs. Houve associação entre as expressões ADRB2 e SLC6A2 com idade, raça, localização do tumor, grau histológico, invasão perineural e status do HPV p16. Foram identificados 898 DEGs entre os grupos. Foi demonstrado que a expressão ADRB2alta / SLC6A2alta influenciou a proliferação, adesão e invasão de células CECPs além da angiogênese. Pacientes com carcinomas espinocelular de laringe e faringe apresentando expressão ADRB2alta / SLC6A2alta tiveram menor sobrevida. Por fim, 56 drogas antineoplásicas e imunoterápicas aprovadas pelo Food Drugs Administration foram identificadas como potenciais alvos para o tratamento personalizado. Significância: Estes achados sugerem fortemente um papel proeminente da sinalização beta-adrenérgica no CECPs ao estimular um fenótipo tumoral mais agressivo. Estas alterações tiveram um impacto negativo no prognóstico dos pacientes com CECP em região de faringe e laringe(AU)

Chronic stress leads to the activation of the beta-adrenergic pathway. Its activation has been implicated in the progression of different types of cancer but its role on head and neck squamous cell carcinomas (HNSCCs) remains undefined. The aim of this study was to investigate the influence of the beta-adrenergic pathway activation in the progression of HNSCCs, assess its impact in the survival of the patients, and explore the potential targets. Five hundred and twenty The Cancer Genome Altas patients with primary HNSCCs were divided in two groups: ADRB2low / SLC6A2low and ADRB2high / SLC6A2high. The association of clinicopathological and genomic features between the groups was analyzed using a bioinformatic approach. Differentially expressed genes (DEGs) were identified through differential expression analysis. Survival analysis was also performed based on ADRB2 and SLC6A2 expressions. Potential drugs for treatment of HNSCC were identified based on the DEGs. There was association between ADRB2 and SLC6A2 expressions with age, race, tumor site, histologic grade, perineural invasion, and HPV p16 status. It was identified 898 DEGs between the groups. It was demonstrated that ADRB2high / SLC6A2high expression influenced HNSCC cells proliferation, adhesion, invasion, and angiogenesis. Patients with larynx and pharynx squamous cell carcinomas presenting ADRB2high / SLC6A2high expression showed had lower survival rates. Finally, 56 Food Drugs Administration-approved antineoplastic and immunotherapeutic drugs were identified as potential targets for the personalized treatment. Significance: These findings strongly suggest a prominent role of beta-adrenergic pathway in HNSCC by stimulating a more aggressive tumoral phenotype. These alterations were shown to negatively impact the prognosis of patients with larynx and pharynx squamous cell carcinomas(AU)

Transhiatal esophagectomy in squamous cell carcinoma of the esophagus: what are the best indications? / Esofagectomia transhiatal no carcinoma de células escamosas do esôfago: quais são suas melhores indicações?

ABSTRACT Background: Southern Brazil has one of the highest incidences of esophageal squamous cell carcinoma in the world. Transthoracic esophagectomy allows more complete abdominal and thoracic lymphadenectomy than transhiatal. However, this one is associated with less morbidity. Aim: To analyze the outcomes and prognostic factors of squamous esophageal cancer treated with transhiatal procedure. Methods: All patients selected for transhiatal approach were included as a potentially curative treatment and overall survival, operative time, lymph node analysis and use of neoadjuvant therapy were analyzed. Results: A total of 96 patients were evaluated. The overall 5-year survival was 41.2%. Multivariate analysis showed that operative time and presence of positive lymph nodes were both associated with a worse outcome, while neoadjuvant therapy was associated with better outcome. The negative lymph-node group had a 5-year survival rate of 50.2%. Conclusion: Transhiatal esophagectomy can be safely used in patients with malnutrition degree that allows the procedure, in those with associated respiratory disorders and in the elderly. It provides considerable long-term survival, especially in the absence of metastases to local lymph nodes. The wider use of neoadjuvant therapy has the potential to further increase long-term survival.

RESUMO Racional: O sul do Brasil tem uma das maiores incidências de carcinoma epidermoide do esôfago no mundo. A esofagectomia transtorácica permite linfadenectomia abdominal e torácica mais completa do que a transhiatal. No entanto, esta está associado à menor morbidade. Objetivo: Analisar os desfechos e fatores prognósticos do câncer epidermoide do esôfago que foram tratados com procedimento transhiatal. Métodos: Foram incluídos todos os pacientes selecionados para abordagem transhiatal como tratamento potencialmente curativo correlacionando sobrevida geral, tempo operatório, análise de linfonodos e uso de terapia neoadjuvante. Resultados: Foram avaliados 96 pacientes. A sobrevida geral em cinco anos foi de 41,2%. A análise multivariada mostrou que o tempo operatório e a presença de linfonodos positivos foram associados a pior resultado, enquanto a terapia neoadjuvante contribuiu para melhor resultado. O grupo de linfonodos negativos teve taxa de sobrevivência em cinco anos de 50,2%. Conclusão: A esofagectomia transhiatal pode ser empregada com segurança em pacientes que apresentem desnutrição com grau que permita o procedimento, nos com distúrbios respiratórios associados e nos idosos. Proporciona sobrevida em longo prazo considerável, especialmente na ausência de metástases para linfonodos locais. O uso mais amplo da terapia neoadjuvante tem o potencial de aumentar ainda mais a sobrevida em longo prazo.