Correlação da imunoexpressão do fator de choque térmico 1 (hsf1) com aspectos clinicopatológicos em carcinomas de células escamosas de língua oral

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.

Transformation to "high grade" neuroendocrine carcinoma as an acquired drug resistance mechanism in EGFR-mutant lung adenocarcinoma.

Several different acquired resistance mechanisms of EGFR mutant lung adenocarcinoma to EGFR-tyrosine kinase inhibitor (TKI) therapy have been described, most recently transformation to small cell lung carcinoma (SCLC). We describe the case of a 46-year-old female with relapsed EGFR exon 19 deletion lung adenocarcinoma treated with erlotinib, and on resistance, cisplatin-pemetrexed. Liver rebiopsy identified an afatinib-resistant combined SCLC and non-small cell carcinoma with neuroendocrine morphology, retaining the EGFR exon 19 deletion. This case highlights acquired EGFR-TKI resistance through transformation to the high-grade neuroendocrine carcinoma spectrum and that that such transformation may not be evident at time of progression on TKI therapy.

Giant condyloma acuminatum quickly growing. Case report

Background. Giant Condyloma Acuminatum (GCA) is a rare, slow growing, large cauliflower tumor of the penile foreskin and perianal region with benign histologic appearance but high propensity for local invasion and recurrences. GCA is associated with Human Papilloma Virus (HPV) types 6 and 11 and it also has considerable risk of neoplastic transformation into fully invasive squamous cell carcinoma into about 5 years. Objective. Because of the rarity of perianal GCA, to date there is no general agreement on the best method for treatment. We wanted to know if surgical approach only was a good method to treat our case. Case report. A 28 years old man, HIV-negative, with a 4 years history of perianal GCA quickly growing underwent full tickness local excision at least 0,7 cm margin of normal tissue with skin grafting taken from the thighs. Fecal contamination was avoided by diet and loperamide per os. At two years follow-up no recurrence was detected. Conclusion. Surgical approach with full tickness excision and immediate skin-grafting and regular follow-up demonstrated effective to treat GCA and to minimize disease recurrence.

Revisão de literatura de Leucoplasia Oral

Similar a outras lesões de pele, têm sido identificadas lesões precursoras de carcinoma de células escamosas na mucosa da cavidade oral. Na boca, apresenta-se frequentemente em forma de placa branca, denominada leucoplasia. Na conferência de 2005, a leucoplasia foi definida pela OMS como “uma placa ou mancha branca que não pode ser caracterizada clínica ou patologicamente como qualquer outra doença.” Leucoplasia é, portanto, um diagnóstico clínico de exclusão. A frequência de apresentar displasia epitelial, carcinoma in situ, carcinoma verrucoso ou carcinoma de células de escomosas invasivo na leucoplasia oral varia de 8,6% a 60%. A transformação maligna anual de leucoplasia é de 1% a 5 %. Sendo assim, é de fundamental importância, por parte dos profissionais na saúde e principalmente dos médicos dentistas terem conhecimento da leucoplasia oral para que possam suspeitar, fazer o diagnóstico ou encaminhar a profissionais competentes precocemente para o manejamento dessas lesões.

Molecular and biochemical mechanisms involved in the toxicity of a marine cyanobacteria compound on cancer cell lines

In recent years marine biotechnology has revealed a crucial role in the future of bioindustry. Among the many marine resources, cyanobacteria have shown great potential in the production of bioactive compounds with diverse applicability. The pharmacological potential of these organisms has been one of the most explored areas in particular its antibacterial, antifungal and anticancer potential. This work was based on the assessment of potential anticancer compound E13010 F 5.4 isolated from marine cyanobacteria strain Synechocystis salina LEGE 06099. Thus the aim of this work was to explore molecular and biochemical mechanisms underlying the bioactivity detected in human cancer cells, specifically in lines RKO colon carcinoma and HT-29. The isolation of the compound was performed from biomass obtained by large-scale culture. To obtain the compound fractionation was carried and confirmation and isolation performed by Nuclear Magnetic Resonance (NMR), Thin Layer Chromatography (TLC) and High-Performance Liquid Chromatography (HPLC). Cell viability assays were performed based on reduction of 3- (4,5-dimetiltiaziol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) to assess the cytotoxic potential of the compound. From the battery of cell lines RKO (colon carcinoma), HT-29 (colorectal adenocarcinoma), MG-63 (osteosarcoma) and T47D (breast carcinoma) the cell lines RKO and HT-29 were selected for elucidation of mechanisms of cytotoxicity. For the elucidation of the mechanisms involved in cytotoxicity the cell lines RKO and HT29 were exposed to the compound. A genomic approach based in the mRNA expression of genes involved in apoptosis and cell cycle by Real-Time PCR and a proteomic approach based on the separation of proteins by two-dimensional electrophoresis (2DGE) was performed. For mRNA expression were selected the genes RPL8, HPRT1, VDAC, SHMT2, CCNE, CCNB1, P21CIP, BCL-2 and BAD and for proteomics isoelectric focussing between 3 – 10 and molecular weight of 19 – 117 kDa separated by polyacrylamide gels (2DGE). The MTT results confirmed the reduction of the cell viability. The RT-PCR results for the expression of genes studied were not yet fully elucidative. For the cell line RKO there was a significant reduction in the expression of the gene P21CIP, and a tendency for reduction in the BAD gene expression and for increased expression of gene CCNB1, pointing to an effort for cell proliferation. In HT-29 cell line, there was a tendency for increase in the expression of P21CIP and BAD, which may explain the reduction in cell viability. The 2DGE results indicate proteomic patterns with differentially altered spots in the treated and control cells with both qualitative and quantitative differences, and differences in response between the RKO and HT-29 cell lines.

Biopsy case mix and diagnostic yield at a Malawian central hospital

Cancer is a major disease burden worldwide resulting in high morbidity and mortality. It is the leading cause of mortality in developed countries and is one of the three leading causes of death for adults in developing countries. Pathological examination of tissue biopsies with histological confirmation of a correct cancer diagnosis is central to cancer care. Without an accurate and specific pathologic diagnosis, effective treatment cannot be planned or delivered. In addition, there are marked geographical variations in incidence of cancer overall, and of the specific cancers seen. Much of the published literature on cancer incidence in developing countries reflects gross estimates and may not reflect reality. Performing baseline studies to understand these distributions lays the groundwork for further research in this area of cancer epidemiology. Our current study surveys and ranks cancer diagnoses by individual anatomical site at Queen Elizabeth Central Hospital (QECH) which is the largest teaching and referral hospital in Malawi. A retrospective study was conducted reviewing available pathology reports over a period of one full year from January 2010 to December 2010 for biopsies from patients suspected clinically of having cancer. There were 544 biopsies of suspected cancer, taken from 96 anatomical sites. The oesophagus was the most common biopsied site followed by breast, bladder, bone, prostate, bowel, and cervical lymph node. Malignancies were found in biopsies of the oesophagus biopsies (squamous cell carcinoma, 65.1%; adenocarcinoma, 11.6%), breast (57.5%), bladder (squamous cell carcinoma, 53.1%) and stomach (37.6%). Our study demonstrates that the yield of biopsy for clinically suspected malignancy was greater than 50% for the 11 most common sites and provides a current survey of cancer types by site present in the population reporting to our hospital.

Environmental risk factors for oesophageal cancer in Malawi: A case-control study

Aim There is a high burden of oesophageal cancer in Malawi with dismal outcomes. It is not known whether environmental factors are associated with oesophageal cancer. Without knowing this critical information, prevention interventions are not possible. The purpose of this analysis was to explore environmental factors associated with oesophageal cancer in the Malawian context. Methods A hospital-based case-control study of the association between environmental risk factors and oesophageal cancer was conducted at Kamuzu Central Hospital in Lilongwe, Malawi and Queen Elizabeth Central Hospital in Blantyre, Malawi. Ninety-six persons with squamous cell carcinoma and 180 controls were enrolled and analyzed. These two groups were compared for a range of environmental risk factors, using logistic regression models. Unadjusted and adjusted odds ratios and 95% confidence intervals (CI) were calculated. Results Firewood cooking, cigarette smoking, and use of white maize flour all had strong associations with squamous cell carcinoma of the oesophagus, with adjusted odds ratios of 12.6 (95% CI: 4.2-37.7), 5.4 (95% CI: 2.0-15.2) and 6.6 (95% CI: 2.3-19.3), respectively. Conclusions Several modifiable risk factors were found to be strongly associated with squamous cell carcinoma. Research is needed to confirm these associations and then determine how to intervene on these modifiable risk factors in the Malawian context.

Síntese verde de nanopartículas de prata a partir de extrato aquoso do tubérculo de Curcuma longa associadas à quitosana e avaliação da atividade antitumoral in vitro em câncer de pele não melanoma (linhagem A431)

Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Biológicas, Programa de Pós-Graduação Nanociência e Nanobiotecnologia, 2016.

Imunoterapia

O cancro oral é uma neoplasia maligna relativamente frequente, sendo por isso responsável por uma taxa de mortalidade elevada. Em particular, o carcinoma espinocelular é o tipo histológico mais frequente das neoplasias malignas da cavidade oral, estando claramente associada a factores de risco como o tabaco, o consumo de álcool e a infecção pelo vírus do papiloma humano (HPV). Actualmente, no mundo ocidental, observa-se um aumento na incidência do cancro da língua que parece estar relacionado com infecções pelos vírus HPV. Tendo em conta os fenómenos associados à cancerização da mucosa oral e a progressão do mesmo, este trabalho tem como função a pesquisa de possíveis alternativas de tratamentos, nomeadamente a imunoterapia, com a utilização de anticorpos monoclonais, terapia de vacinas, terapia de transferência adoptiva de células T, entre outras, uma vez que nem sempre os tratamentos convencionais como a quimioterapia, radioterapia, ou tratamento cirúrgico se revelam completamente eficazes. Contudo, existe uma carência de protocolos definidos, sendo a imunoterapia ainda uma terapêutica a evoluir, por isso esta monografia pretende fazer uma revisão sobre o ‘’estado da arte’’ deste tema tão complexo, com base em literatura de vários autores ao longo desta última década. Este trabalho pretende mencionar novos alvos terapêuticos que permitem desenhar terapêuticas mais dirigidas e, eventualmente, com menos efeitos adversos. A utilização por exemplo do cetuximab (anti-EGFR), que na prática clínica é já uma realidade.

Avaliação do nível de conhecimento geral e grau de alerta sobre cancro oral numa população do nordeste transmontano

O cancro é um dos principais causadores de milhões de mortes em todo o mundo e sendo o cancro oral, especificamente, a sexta neoplasia mais frequente a nível mundial. Todos os anos são diagnosticados mais de 500 mil novos casos, sendo que as altas taxas de mortalidade e mortalidade não se têm alterado ao longo dos anos. A maior incidência de cancro oral encontra-se na Ásia e na Europa do Sul. Em Portugal, mais precisamente em 2012, foram diagnosticados cerca de 1924 novos casos de cancro oral, dos quais 967 ocorreram em homens. O carcinoma espinocelular é o tipo histológico mais comum, sendo que 90% dos casos de cancro oral são deste tipo. Sabe-se também que esta variante é mais frequente no sexo masculino entre a 5ª e 6 ª década de vida apesar de, a incidência no sexo feminino, ter vindo a aumentar, devido à contínua exposição ao tabaco, álcool e a outros factores de risco. Como foi dito anteriormente, o cancro oral tem uma alta taxa de mortalidade e de morbilidade e, apesar dos avanços no diagnóstico, no tratamento e no conhecimento de quais os factores de risco desta patologia, a taxa de sobrevivência ainda é inferior a 50% o que revela que, o grande problema, passa pelo diagnóstico do cancro em estádios avançados. Assume-se então que, grande parte dos casos de cancro oral, poderiam ter sido evitados se houvesse maior conhecimento e grau de alerta sobre a doença o que tendencialmente, levaria a diagnósticos mais precoces. Neste sentido, este estudo tem como propósito a avaliação do nível de conhecimento geral e do grau de alerta de uma população do interior do país, mais precisamente do Nordeste Transmontano, bem como, efectuar o registo da percepção dos inquiridos relativamente a esta patologia, passando pelo reconhecimento da doença, pelo conhecimento epidemiológico e etiológico, e pela melhor percepção a nível de sinais e sintomas clínicos próprios desta patologia.

Tecniche in vivo ancillari alla chirurgia di Mohs: ruolo delle nuove tecnologie nella valutazione dei margini chirurgici delle neoplasie cutanee maligne

Le neoplasie cutanee di tipo non-melanoma (non-melanoma skin cancers, NMSCs), quali il carcinoma a cellule basali (basal cell carcinoma, BCC) e il carcinoma a cellule squamose (squamous cell carcinoma, SCC) possono mostrare invasività locale e alto tasso di recidiva. La chirurgia microscopicamente controllata di Mohs (Mohs micrographic surgery, MMS) permette di eseguire una valutazione istologica immediata dei margini chirurgici delle neoplasie contestualmente alla loro escissione. Nel nostro studio abbiamo valutato del ruolo delle tecnologie in vivo (dermatoscopia e microscopia confocale a riflettanza, MCR) nella definizione dei margini preoperatori di NMSC ad alto rischio del volto e descritto la nostra esperienza con chirurgia tradizionale e MMS. Sono stati valutati 234 pazienti operati nel triennio 2019-2021: 39 con MMS e guida videodermatoscopica (Gruppo 1) e 195 con chirurgia tradizionale e guida videodermatoscopica (Gruppo 2). I pazienti operati nel periodo 2013-2018 (con MMS, Gruppo 3 (n = 241), e con chirurgia tradizionale, Gruppo 4 (n = 1086)) sono stati usati come confronto. La radicalità chirurgica è stata ottenuta nel Gruppo 1 nel 92,3% dei casi, con 1,2 steps in media di MMS (versus 1,7 nel Gruppo 3), nel Gruppo 2 nell’84,5% dei casi. La percentuale di non radicalità è stata: 7,7% nel Gruppo 1, 15,9% nel Gruppo 2, 6,2% nel Gruppo 3, 17,9% nel Gruppo 4. I tassi di recidiva sono stati: 5,1% nel Gruppo 1, 3,6% nel Gruppo 2, 4,1% nel Gruppo 3, 5,9% nel Gruppo 4, soprattutto in BCC di tipo sclerodermiforme e infiltrante. La MCR prechirurgica è stata utilizzata in 11 pazienti, con alcuni limiti nel delineare BCC di tipo sclerodermiforme e infiltrante. In conclusione, la videodermatoscopia e la MCR appaiono valide tecniche ancillari alla MMS e anche alla chirurgia tradizionale nel trattamento dei NMSCs. La MMS appare indicata soprattutto nei pazienti giovani e salvaguarda l’outcome estetico e funzionale.

Identification of potential miRNAs related to classification and treatment response of actinic keratosis

The Workflow activity was the following: Preliminary phase: Identification of 18 Formalin-fixed paraffin embedded (FFPE) samples (9 patients) («matched» 9 AK lesions and 9 SCC lesions). Working on biopsies samples we perform an extraction and RNA analysis with droplet Digital PCR (ddPCR) and we perform the data analysis. Second and final step phase: Evaluation of additional 39 subjects (36 men and 3 women). Results: We perform an evaluation and comparison of the following miRNA: miR-320 (a miRNA involved in apoptosis and cell proliferation control; miR-204, a miRNA involved in cell proliferation in and miRNA-16-5p, a miRNA involved in apoptosis).Conclusion: Our data suggest that there is no significant variation in the expression of the three tested microRNAs between adjacent AK lesions and squamous-cell carcinoma. However, a relevant trend has been observed Furthermore, by evaluating the miRNA expression trend between keratosis and carcinoma of the same patient, it is observed that there is no "uniform trend": for some samples the expression rises for the transition from AK to SCC and viceversa.

A Putative Otu Domain-containing Protein 1 Deubiquitinating Enzyme Is Differentially Expressed In Thyroid Cancer And Identifies Less-aggressive Tumours.

This study aimed to identify novel biomarkers for thyroid carcinoma diagnosis and prognosis. We have constructed a human single-chain variable fragment (scFv) antibody library that was selected against tumour thyroid cells using the BRASIL method (biopanning and rapid analysis of selective interactive ligands) and phage display technology. One highly reactive clone, scFv-C1, with specific binding to papillary thyroid tumour proteins was confirmed by ELISA, which was further tested against a tissue microarray that comprised of 229 thyroid tissues, including: 110 carcinomas (38 papillary thyroid carcinomas (PTCs), 42 follicular carcinomas, 30 follicular variants of PTC), 18 normal thyroid tissues, 49 nodular goitres (NG) and 52 follicular adenomas. The scFv-C1 was able to distinguish carcinomas from benign lesions (P=0.0001) and reacted preferentially against T1 and T2 tumour stages (P=0.0108). We have further identified an OTU domain-containing protein 1, DUBA-7 deubiquitinating enzyme as the scFv-binding antigen using two-dimensional polyacrylamide gel electrophoresis and mass spectrometry. The strategy of screening and identifying a cell-surface-binding antibody against thyroid tissues was highly effective and resulted in a useful biomarker that recognises malignancy among thyroid nodules and may help identify lower-risk cases that can benefit from less-aggressive management.

Evolução dos doentes com citologia oncótica alterada e colposcopia anal normal

A citologia anal vem sendo usada para rastreamento do carcinoma anal e suas lesões precursoras nas populações de risco. Quando o raspado do canal anal mostra alterações citológicas está indicada o exame com colposcópio e ácido acético para identificar e realizar biópsia para confirmar o achado. Poucos estudos mostram o seguimento dos doentes tratados de condilomas acuminados perianais. Temos usado os métodos em associação e encontrado lesões subclínicas em metade dos doentes, cujo exame proctológico não revelava doença HPV induzida. Essas lesões são tratadas com tópicos. Entretanto, algumas citologias estavam alteradas e a colposcopia anal não revelou doença HPV induzida. O objetivo deste estudo foi observar o comportamento dessas lesões no seguimento semestral, durante 12 meses, e avaliar se a periodicidade da reavaliação foi suficiente para evitar o aparecimento das lesões de alto grau ou superior. Encontramos 58 (21%) entre 273 doentes nessas condições. As reavaliações de 22 deles após um ano mostraram que as colposcopias permaneceram normais em 17 (74%), sendo que em cinco (22%) a citologia voltou aos padrões normais e 12 (52%) persistiram com alterações. Os outros seis (26%) desenvolveram lesões clínicas ou subclínicas provocadas pelo HPV. As contagens de linfócitos T CD4 dos doentes HIV-positivos foram inferiores nos doentes cujas lesões progrediram. Os resultados permitiram concluir que as alterações podem progredir ou regredir neste grupo distinto de doentes, sendo relacionada à imunidade, e que o intervalo de seis meses é suficiente para cada reavaliação.

Characterization of global transcription profile of normal and HPV-immortalized keratinocytes and their response to TNF treatment

Background: Persistent infection by high risk HPV types (e.g. HPV-16, -18, -31, and -45) is the main risk factor for development of cervical intraepithelial neoplasia and cervical cancer. Tumor necrosis factor (TNF) is a key mediator of epithelial cell inflammatory response and exerts a potent cytostatic effect on normal or HPV16, but not on HPV18 immortalized keratinocytes. Moreover, several cervical carcinoma-derived cell lines are resistant to TNF anti-proliferative effect suggesting that the acquisition of TNF-resistance may constitute an important step in HPV-mediated carcinogenesis. In the present study, we compared the gene expression profiles of normal and HPV16 or 18 immortalized human keratinocytes before and after treatment with TNF for 3 or 60 hours. Methods: In this study, we determined the transcriptional changes 3 and 60 hours after TNF treatment of normal, HPV16 and HPV18 immortalized keratinocytes by microarray analysis. The expression pattern of two genes observed by microarray was confirmed by Northern Blot. NF-kappa B activation was also determined by electrophoretic mobility shift assay (EMSA) using specific oligonucleotides and nuclear protein extracts. Results: We observed the differential expression of a common set of genes in two TNF-sensitive cell lines that differs from those modulated in TNF-resistant ones. This information was used to define genes whose differential expression could be associated with the differential response to TNF, such as: KLK7 (kallikrein 7), SOD2 (superoxide dismutase 2), 100P (S100 calcium binding protein P), PI3 (protease inhibitor 3, skin-derived), CSTA (cystatin A), RARRES1 (retinoic acid receptor responder 1), and LXN (latexin). The differential expression of the KLK7 and SOD2 transcripts was confirmed by Northern blot. Moreover, we observed that SOD2 expression correlates with the differential NF-kappa B activation exhibited by TNF-sensitive and TNF-resistant cells. Conclusion: This is the first in depth analysis of the differential effect of TNF on normal and HPV16 or HPV18 immortalized keratinocytes. Our findings may be useful for the identification of genes involved in TNF resistance acquisition and candidate genes which deregulated expression may be associated with cervical disease establishment and/or progression.