The Complexity of Tobacco Smoke-Induced Mutagenesis in Head and Neck Cancer.

Tobacco smoke, alone or combined with alcohol, is the predominant cause of head and neck cancer (HNC). Here, we further explore how tobacco exposure contributes to cancer development by mutational signature analysis of 265 whole-genome sequenced HNC from eight countries. Six tobacco-associated mutational signatures were detected, including some not previously reported. Differences in HNC incidence between countries corresponded with differences in mutation burdens of tobacco-associated signatures, consistent with the dominant role of tobacco in HNC causation. Differences were found in the burden of tobacco-associated signatures between anatomical subsites, suggesting that tissue-specific factors modulate mutagenesis. We identified an association between tobacco smoking and three additional alcohol-related signatures indicating synergism between the two exposures. Tobacco smoking was associated with differences in the mutational spectra and repertoire of driver mutations in cancer genes, and in patterns of copy number change. Together, the results demonstrate the multiple pathways by which tobacco smoke can influence the evolution of cancer cell clones.

Changes in Body Mass Index Are Associated with Squamous Cell Carcinomas of Oral Cavity, Oropharynx and Larynx: A Case-Control Study in Brazil.

Head and neck cancer (HNC) significantly impacts nutritional status because the tumor limits swallowing function. In this sense, it is important to monitor the nutritional status throughout the life of any individual. A multicenter case-control study was carried out to analyze the BMI at 30 years of age, two years before diagnosis and at the time of diagnosis of individuals with oral cavity, oropharynx, and larynx cancers. It was observed that a 5% reduction in BMI during the two years before enrollment was associated with an increased risk of the oral cavity (OR = 3.73), oropharyngeal OR = 5.25), and laryngeal (OR = 5.22). Reduced BMI of more than 5% over two years before diagnosis was associated with HNC. Weight loss remained significant at diagnosis, but it is not possible to exclude reverse causality since most cases are at an advanced stage. BMI monitoring of individuals at potential risk for HNC can promote early diagnosis and nutritional interventions for HNC.

Risk factors associated with head and neck cancer in former smokers: A Brazilian multicentric study.

BACKGROUND: Reduced tobacco consumption in the population has not been associated with reduced incidence rates of head and neck cancer in several countries. OBJECTIVE: To explore the associations between HNC and sociodemographic characteristics and lifestyle of former smokers from three Brazilian cancer centers. METHODS: A multicenter case-control study was conducted with 229 former smokers diagnosed with squamous cell carcinoma of the oral cavity, oropharynx, larynx, and 318 controls (former smokers without head and neck cancer). Bivariate and multiple logistic regression analyses were conducted to estimate odds ratios (ORs) with a 95% confidence interval (CI). RESULTS: 11-20 years after smoking cessation showed significant impact on HNC reduction (OR 0.22, 95% CI, 0.12-0.39), which reached 82% (95% CI, 0.09-0.35) among 20 + former smokers when compared to individuals who had stopped smoking for up to 5 years. A history of high-intensity smoking (>40 pack-years) increased HNC risk by 2.09 times (95% CI 1.13-3.89) when compared to subjects who smoked up to 20 pack-years. Past alcohol consumption (OR 1.99, 95% CI, 1.06-3.82) was also associated with head and neck cancer risk in former smokers when compared to no alcohol consumption. There was a decreased head and neck cancer risk in former smokers who had high school level of education (OR 0.38, 95% CI, 0.16-0.91) compared to illiterate former smokers; and former smokers with moderate intake of vegetables (OR 0.49, 95% CI, 0.28-0.85) and fruits (OR 0.43, 95% CI, 0.25-0.73) compared to those with low intake. CONCLUSION: Head and neck cancer risk in former smokers decreases after 11 years after smoking cessation, former smokers with past alcohol consumption showed an increased risk of HNC. High school level of education and moderate intake of vegetables and fruits reduced HNC risk among former smokers.

Differential Expression of Potential Biomarkers of Oral Squamous Cell Carcinoma Development.

To evaluate molecular epithelial changes, we investigated whether a profile of survivin, cyclin dependent kinase inhibitor 2A (CDKN2A), epidermal growth factor receptor (EGFR), polo like kinase 1 (PLK1), p63, p40 (Δnp63 isoform), cyclin D1 (CCND1) and BCL2 apoptosis regulator (BCL2) proteins could predict malignant transformation. Different tissue segments (tumor adjacent epithelium; dysplasia and tumor) from a total of 109 patients were analyzed by immunohistochemistry. An increased expression of survivin (p < 0.001), PLK1 (p = 0.001), and p63 (p < 0.001) in parallel to reduced immunostaining of p40 (p < 0.001) and BCL2 (p = 0.029) was observed among the tissue segments analyzed. Our study revealed that survivin, PLK1, p63, p40 and BCL2 play a role in oral tumorigenesis and represent promising biomarkers able to recognize mesenchymal phenotype induction in the transition from nonmalignant cells to tumor cells. These results reveals critical interaction between survivin, PLK1, p63, p40 promising proteins during invasive carcinoma development.

Prognostic value of pretreatment cardiovascular biomarkers in head and neck squamous cell carcinoma.

OBJECTIVES: To examine the prognostic significance of pretreatment C-reactive protein (CRP), N-terminal pro-brain natriuretic peptide (NT-proBNP), and cardiac troponin T (cTnT) levels on all-cause mortality 3 years after head and neck squamous cell carcinoma (HNSCC) diagnosis. SUBJECTS AND METHODS: Data from 118 consecutive HNSCC patients, treated between 2012 and 2015, were evaluated prospectively. The impact of CRP, high-sensitive (hs)-cTnT, and NT-proBNP levels on the 3-year overall survival was estimated using the Kaplan-Meier method and Cox proportional hazard models. RESULTS: During the 36-month follow-up, 37 patients (31.35%) died. Multivariate analysis revealed that elevated CRP (Hazard ratio: 3.71, 95% CI: 1.44-9.53, p = .007) and NT-proBNP levels (Hazard ratio: 5.04, 95% CI: 2.02-12.55, p = .001) were associated with negative prognosis, independent on age, sex, smoking and alcohol status, TNM classification, tumor site, body mass index (BMI), systolic blood pressure (SBP), and treatment modality (except for radiotherapy). hs-cTnT had no influence over the prognosis, but it was correlated with TNM classification and SBP. CRP was significantly correlated with BMI and TNM classification, and NT-proBNP with SBP and hs-cTnT. CONCLUSIONS: Pretreatment CRP and NT-proBNP levels were identified as independent prognostic markers for poor clinical outcome 3 years after HNSCC diagnosis.

Hypermethylation status of DAPK, MGMT and RUNX3 in HPV negative oral and oropharyngeal squamous cell carcinoma.

Squamous cell carcinoma of the oral cavity and oropharynx is the sixth most common type of cancer in the world. During tumorigenesis, gene promoter hypermethylation is considered an important mechanism of transcription silencing of tumor suppressor genes, such as DAPK, MGMT and RUNX3. These genes participate in signaling pathways related to apoptosis, DNA repair and proliferation whose loss of expression is possibly associated with cancer development and progression. In order to investigate associations between hypermethylation and clinicopathological and prognostic parameters, promoter methylation was evaluated in 72 HPV negative oral and oropharyngeal tumors using methylation-specific PCR. Hypermethylation frequencies found for DAPK, MGMT and RUNX3 were 38.88%, 19.44% and 1.38% respectively. Patients with MGMT hypermethylation had a better 2-year overall survival compared to patients without methylation. Being MGMT a repair gene for alkylating agents, it could be a biomarker of treatment response for patients who are candidates for cisplatin chemotherapy, predicting drug resistance. In view of the considerable levels of hypermethylation in cancer cells and, for MGMT, its prognostic relevance, DAPK and MGMT show potential as epigenetic markers, in a way that additional studies may test its viability and efficacy in clinical management.

Human Papillomavirus E6/E7 mRNA detection by in situ hybridization in oral cavity squamous cell carcinoma.

OBJECTIVE: The aim of this study was to evaluate the application of in situ hybridization using E6/E7 mRNA probes to identify the frequency of high-risk HPV transcriptionally active and the use of HPV status as a prognostic biomarker in oral cavity squamous cell carcinoma (OCSCC). METHODS: Ninety-nine OCSCC samples were evaluated from Hospital Santa Rita de Cassia, Hospital Universitário Cassiano Antônio de Moraes and University Hospitals Coventry and Warwickshire NHS Trust. After tissue microarray construction, the slides were submitted to an in situ hybridization detection method for HPV E6/E7 mRNA. HPV status was designated a binary classification. Multiple logistic regression examined the association of HPV with clinical features and other risk factors, using SPSS® software. For all hypothesis tests, a significance level of p ≤ 0.05 was considered. RESULTS: HPV frequency in oral squamous cell carcinoma was 8%. There was no association between HPV and clinical variables and between the main prognostic features and known risk factors. There was no difference in the prevalence of HPV for oral cavity squamous cell carcinoma by geography (Brazil vs UK). CONCLUSIONS: A low frequency of E6/E7 mRNA by RNA in situ hybridization was found in oral cavity squamous cell carcinoma, which supports the evidence that HPV-driven cancer of the oral cavity is uncommon.

Predictors of Survival After Head and Neck Squamous Cell Carcinoma in South America: The InterCHANGE Study.

PURPOSE: Head and neck squamous cell carcinoma (HNSCC) incidence is high in South America, where recent data on survival are sparse. We investigated the main predictors of HNSCC survival in Brazil, Argentina, Uruguay, and Colombia. METHODS: Sociodemographic and lifestyle information was obtained from standardized interviews, and clinicopathologic data were extracted from medical records and pathologic reports. The Kaplan-Meier method and Cox regression were used for statistical analyses. RESULTS: Of 1,463 patients, 378 had a larynx cancer (LC), 78 hypopharynx cancer (HC), 599 oral cavity cancer (OC), and 408 oropharynx cancer (OPC). Most patients (55.5%) were diagnosed with stage IV disease, ranging from 47.6% for LC to 70.8% for OPC. Three-year survival rates were 56.0% for LC, 54.7% for OC, 48.0% for OPC, and 37.8% for HC. In multivariable models, patients with stage IV disease had approximately 7.6 (LC/HC), 11.7 (OC), and 3.5 (OPC) times higher mortality than patients with stage I disease. Current and former drinkers with LC or HC had approximately 2 times higher mortality than never-drinkers. In addition, older age at diagnosis was independently associated with worse survival for all sites. In a subset analysis of 198 patients with OPC with available human papillomavirus (HPV) type 16 data, those with HPV-unrelated OPC had a significantly worse 3-year survival compared with those with HPV-related OPC (44.6% v 75.6%, respectively), corresponding to a 3.4 times higher mortality. CONCLUSION: Late stage at diagnosis was the strongest predictor of lower HNSCC survival. Early cancer detection and reduction of harmful alcohol use are fundamental to decrease the high burden of HNSCC in South America.

Consumption of minimally processed foods as protective factors in the genesis of squamous cell carcinoma of the head and neck in Brazil.

BACKGROUND: Head and neck cancer (HNC) is the sixth most common cancer, and two-fifths of cases could be avoided by changing lifestyle and eating habits. METHODS: This multicenter case-control study was conducted under the International Consortium on Head and Neck Cancer and Genetic Epidemiology, coordinated by the International Agency for Research on Cancer. This consortium evaluated associations between minimally processed food consumption and the risk of HNC in three Brazilian states. RESULTS: We evaluated 1740 subjects (847 cases and 893 controls). In multiple analyses including recognized risk factors for HNC, the consumption of apples and pears was associated with reduced risks of oral cavity and laryngeal cancers; the consumption of citrus fruits and fresh tomatoes was associated with a reduced risk of oral cavity cancer; the consumption of bananas was associated with a reduced risk of oropharynx cancer; the consumption of broccoli, cabbage, and collard greens was associated with reduced risks of laryngeal and hypopharyngeal cancers; and the consumption of carrots and fresh fruits was associated with a reduced risk of hypopharyngeal cancer. CONCLUSIONS: The consumption of a heathy diet rich in fruits and vegetables was associated with a reduced risk of HNC. Public policies, including government subsidies, are essential to facilitate logistical and financial access to minimally processed foods, thereby strengthening environments that promote healthy behavior.

Frequency of HPV in oral cavity squamous cell carcinoma.

BACKGROUND: The prevalence of high-risk human papillomavirus (HPV) DNA in cases of oral cavity squamous cell carcinoma (SCC) varies widely. The aim of this study is to investigate the frequency of high-risk HPV DNA in a large Brazilian cohort of patients with oral cavity SCC. METHODS: Biopsy and resected frozen and formalin-fixed paraffin-embedded specimens of oral cavity SCC were available from 101 patients who were recruited at two Brazilian centres. Stringent measures with respect to case selection and prevention of sample contamination were adopted to ensure reliability of the data. Nested PCR using MY09/MY11 and GP5+/GP6+ as well as PGMY09/11 L1 consensus primers were performed to investigate the presence of HPV DNA in the tumours. HPV-positive cases were subjected to direct sequencing. Shapiro-Wilk and Student t test were used to evaluate data normality and to compare the means, respectively. Qualitative variables were analysed by logistic regression. RESULTS: Our results demonstrate that the frequency of high-risk HPV types in oral cavity SCC is very low and is less than 4%. All HPV-positive cases were HPV16. In addition, our results do not show a significant association between the tumour clinical features and the risk factors (tobacco, alcohol and HPV) for oral cavity SCC. CONCLUSION: In the current study, we observed an overlapping pattern of risk factors that are related to tumour development. This, along with a low frequency of high-risk HPV DNA, supports the findings that HPV is not involved in the genesis of oral cavity SCC in Brazilian population.

Polymorphisms in methylenetetrahydrofolate reductase and cystathionine beta-synthase in oral cancer - a case-control study in southeastern Brazilians / Polimorfismos em metilenotetrahidrofolato redutase, cistationina beta-sintase no câncer de boca - um estudo de caso-controle no Sudeste brasileiro

ABSTRACT INTRODUCTION: Oral squamous cell carcinoma (OSCC) is a serious public health problem, due to its high mortality rate and worldwide rising incidence. OSCC susceptibility is mediated by interactions between genetic and environmental factors. Studies suggest that genetic variants encoding enzymes involved in folate metabolism may modulate OSCC risk by altering DNA synthesis/repair and methylation process. OBJECTIVE: The goals of this study were to evaluate the association of three genotypic polymorphism (MTHFR C677T, MTHFR A1298C and CBS 844ins68) and oral cancer risk in southeastern Brazilians and evaluate the interactions between polymorphisms and clinical histopathological parameters. METHODS: This case-control study included 101 cases and 102 controls in the state of Espírito Santo, Brazil. MTHFR genotyping was done by PCR-RFLP (polymerase chain reaction - restriction fragment length polymorphism) and CBS genotyping by PCR (polymerase chain reaction) analysis. RESULTS: MTHFR C677T polymorphism was associated with lymph node involvement. Genotype CT + TT acted as a protective factor. MTHFR A1298C AC + CC genotype was associated with tumor differentiation, and possibly with a better prognosis. In risk analysis, no correlation was observed between genotypes and OSCC. CONCLUSION: We concluded that MTHFR C677T, MTHFR A1298C and CBS 844ins68 polymorphisms were not associated with OSCC risk in southeastern Brazilians; however, we suggest a prognosis effect associated with MTHFR C677T and A1298C polymorphisms in OSCC.

Resumo Introdução: O carcinoma espinocelular oral (CECO) trata-se de um importante problema de saúde pública, devido à elevada taxa de mortalidade e incidência crescente em todo o mundo. A susceptibilidade ao CECO é mediada por interações entre fatores genéticos e ambientais. Estudos sugerem que as variantes genéticas que codificam as enzimas envolvidas no metabolismo do folato podem modular o risco de CECO, alterando a síntese/reparação do DNA e o processo de metilação. Objetivo: Os objetivos deste estudo foram avaliar a associação de três polimorfismos genotípicos (MTHFR C677T, MTHFR A1298C e CBS 844ins68) e o risco de câncer oral em brasileiros da região Sudeste, e avaliar as interações entre polimorfismos e parâmetros clínico-histopatológicos. Método: Este estudo de caso-controle incluiu 101 casos e 102 controles no estado do Espírito Santo, Brasil. A genotipagem do polimorfismo MTHFR foi realizada por PCR-RFLP (Reação de Polimerase em Cadeia - Polimorfismo no Comprimento de Fragmento de Restrição) e a do CBS por análise da PCR (Reação de Polimerase em Cadeia). Resultados: O polimorfismo MTHFR C677T foi associado ao envolvimento de gânglios linfáticos. O genótipo CT + TT atuou como um fator protetor. O genótipo MTHFR A1298C AC + CC foi associado à diferenciação do tumor e, possivelmente, a um prognóstico melhor. Na análise de risco, a correlação entre os genótipos e o CECO não foi observada. Conclusão: Concluímos que os polimorfismos MTHFR C677T, MTHFR A1298C e CBS 844ins68 não estão associados ao risco de CECO nos brasileiros da região Sudeste; no entanto, sugerimos um efeito prognóstico associado aos polimorfismos MTHFR C677T e A1298C em CECO.

Polymorphisms in methylenetetrahydrofolate reductase and cystathionine beta-synthase in oral cancer - a case-control study in southeastern Brazilians.

INTRODUCTION: Oral squamous cell carcinoma (OSCC) is a serious public health problem, due to its high mortality rate and worldwide rising incidence. OSCC susceptibility is mediated by interactions between genetic and environmental factors. Studies suggest that genetic variants encoding enzymes involved in folate metabolism may modulate OSCC risk by altering DNA synthesis/repair and methylation process. OBJECTIVE: The goals of this study were to evaluate the association of three genotypic polymorphism (MTHFR C677T, MTHFR A1298C and CBS 844ins68) and oral cancer risk in southeastern Brazilians and evaluate the interactions between polymorphisms and clinical histopathological parameters. METHODS: This case-control study included 101 cases and 102 controls in the state of Espírito Santo, Brazil. MTHFR genotyping was done by PCR-RFLP (polymerase chain reaction - restriction fragment length polymorphism) and CBS genotyping by PCR (polymerase chain reaction) analysis. RESULTS: MTHFR C677T polymorphism was associated with lymph node involvement. Genotype CT+TT acted as a protective factor. MTHFR A1298C AC+CC genotype was associated with tumor differentiation, and possibly with a better prognosis. In risk analysis, no correlation was observed between genotypes and OSCC. CONCLUSION: We concluded that MTHFR C677T, MTHFR A1298C and CBS 844ins68 polymorphisms were not associated with OSCC risk in southeastern Brazilians; however, we suggest a prognosis effect associated with MTHFR C677T and A1298C polymorphisms in OSCC.

Influence of pain severity on the quality of life in patients with head and neck cancer before antineoplastic therapy.

BACKGROUND: The aim of this study was to assess the severity of pain and its impact on the quality of life (QoL) in untreated patients with head and neck squamous cell carcinoma (HNSCC). METHODS: A study group of 127 patients with HNSCC were interviewed before antineoplastic treatment. The severity of pain was measured using the Brief Pain Inventory (BPI) questionnaire, and the QoL was assessed with the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire Core-30 (EORTC QLQ-C30) and the head and neck module (QLQ-H&N35). RESULTS: The mean age of the patients was 57.9 years, and there was a predominance of men (87.4%). The most frequent site of the primary tumor was the oral cavity (70.6%), and the majority of the patients had advanced cancers (stages III and IV). QoL in early stage of cancer obtained better scores. Conversely, the patients with advanced stage cancer scored significantly higher on the symptom scales regarding fatigue, pain, appetite loss and financial difficulties, indicating greater difficulties. Regard to the severity of pain, patients with moderate-severe pain revealed a significantly worse score than patients without pain. CONCLUSIONS: The severity of pain is statistically related to the advanced stages of cancer and directly affects the QoL. An assessment of the quality of life and symptoms before therapy can direct attention to the most important symptoms, and appropriate interventions can then be directed toward improving QoL outcomes and the response to treatment.

Resumen desayuno de trabajo: "El impacto de los medicamentos de alto costo en oncología" / Working breakfast summary: "The impact of high-cost drugs on oncology"

Los medicamentos de alto costo (MAC) son una realidad preocupante en el tratamiento oncológico actual. El aumento de los precios de las nuevas drogas es dramático y pone en peligro la sustentabilidad del sistema de salud. Con esto, queda demostrado que, si bien la salud es un problema de todos y no tiene precio, lo cierto es que tiene un costo.

Relação entre condição bucal precária e carcinoma de células escamosas oral e orofaríngeo / Relationship between poor oral health and oral and oropharyngeal squamous cell carcinoma

Introdução: Condição bucal precária tem sido relacionada aorisco de desenvolvimento de câncer, no entanto os critérios paradeterminar esta relação ainda não estão bem estabelecidos.Objetivo: O objetivo deste estudo foi avaliar a condição bucale sua relação com o desenvolvimento do carcinoma de célulasescamosas oral e orofaríngeo. Método: Trata-se de estudotransversal descritivo de perfil epidemiológico, onde foram obtidosdados de 150 pacientes sobre gênero, faixa etária, etnia e históriado uso de tabaco e consumo de álcool. A condição bucal foideterminada utilizando os índices de número de dentes perdidos,restaurados e cariados (CPO-D), Perda de Inserção Periodontal(PIP) e Índice Periodontal Comunitário (CPI). Resultados: Aavaliação da condição bucal mostrou que 98,67% dos indivíduosapresentaram grande número de dentes perdidos (média 23,11dentes). Na análise do CPO-D foi observada diferença na faixaetária acima de 50 anos (p< 0.01). Não foram observadasdiferenças significativas relacionadas à quantidade de tabaco ouálcool consumida, bem como ao tempo de consumo de bebidasalcoólicas. O uso do tabaco mostrou relação somente com o índiceCPO-D associado ao tempo de uso superior a 30 anos (p< 0.05).Na análise dos índices CPI e PIP, 74% dos sextantes foramexcluídos devido ao grande número de perdas dentárias. Foramobservados presença de sangramento, cálculo e profundidadede sondagem maior que 4 mm na maioria dos sextantesanalisados. Conclusão: Relação entre condição bucal precária edesenvolvimento de câncer oral e orofaríngeo foi observada nesteestudo, considerando CPO-D, CPI e PIP indicadores relevantespara determinar a condição bucal de pacientes oncológicos.

Promoter hypermethylation in primary squamous cell carcinoma of the oral cavity and oropharynx: a study of a Brazilian cohort.

Epigenetic silencing of cancer-related genes plays an important role in oral/oropharyngeal squamous cell carcinoma (OSCC). We evaluated promoter hypermethylation of 4 cancer-related genes in OSCCs of a Brazilian cohort and determined its relationship with exposure to alcohol, tobacco, HPV infection and clinicopathological parameters. CDKN2A (cyclin-dependent kinase inhibitor 2A or p16), SFN (stratifin or 14-3-3 σ), EDNRB (endothelin receptor B) and RUNX3 (runt-related transcript factor-3) had their methylation patterns evaluated by MSP analysis in OSCC tumors (n = 45). HPV detection was carried out by PCR/RFLP. Aberrant methylation was detected in 44/45 (97.8 %) OSCC; 24.4 % at CDKN2A, 77.8 % at EDNRB, 17.8 % at RUNX3 and 97.8 % at SFN gene. There was no significant association between methylation patterns and clinical parameters. HPV (subtype 16) was detected in 3 out of 45 patients (6 %). Our findings indicate that HPV infection is uncommon and methylation is frequent in Brazilian OSCCs, however, EDNRB and SFN gene methylation are not suitable OSCC biomarkers due to indistinct methylation in tumoral and normal samples. In contrast, CDKN2A and RUNX3 genes could be considered differentially methylated genes and potential tumor markers in OSCCs.