FGF-2 and FGFR-1 immunoexpression in oral tongue squamous cell carcinoma

Orientador: Jacks Jorge JuniorDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: O fator de crescimento fibroblástico 2 (FGF-2) e o receptor do fator de crescimento fibroblástico 1 (FGFR-1) estão associados à maior capacidade de invasão tumoral, proliferação celular, angiogênese e ao potencial de metástase. Este estudo teve como objetivo investigar a expressão de FGF-2 e FGFR-1 na displasia epitelial oral (DEO) e carcinoma espinocelular de língua (CECL). Foram selecionados retrospectivamente cento e sessenta e sete casos, incluindo 85 espécimes cirúrgicos de pacientes com CECL, provenientes do Hospital Onofre Lopes, Natal, Brasil, além de 46 biópsias incisionais de CECL e 36 de DEO provenientes do arquivo do Laboratório de Patologia Oral da Faculdade de Odontologia de Piracicaba (UNICAMP). Cortes de tecido parafinado foram submetidas à reação imunoistoquímica para FGF-2 e FGFR-1. As lâminas foram escaneadas e a marcação imunoistoquímica foi quantificada digitalmente pelo software Aperio Positive Pixel Count v9. Cinco áreas iguais foram selecionadas no estroma e no epitélio tumoral. Os resultados foram exportados e o score final de cada lesão foi obtido através da soma da porcentagem de pixels fracos, moderados e fortes, gerando um valor que variou de 100 a 300. Os casos foram divididos como tendo "baixa expressão" ou "alta expressão" das proteínas de acordo com a mediana dos grupos. FGF-2 e FGFR-1 foram mais expressos em DEO de alto grau do que em DEO de baixo grau, tanto no epitélio, como nas células do estroma (p<0.05). A sobrevivência doença específica (SDE) em 5 anos foi de 47,3% dos pacientes com CECL. A alta expressão de FGF-2 nas células inflamatórias e mesenquimatosas do estroma foi associada à invasão vascular e pior prognóstico. Pacientes com alta expressão de FGF-2 no estroma apresentaram taxa de SDE em 5 anos de 36,7%, contra 59,3% dos pacientes com baixa expressão (HR: 2,272; IC95%: 1.213-4.254; p=0,008). A alta expressão de FGFR-1 no estroma foi correlacionada com metástases linfonodais e metástases à distância. Pacientes com alta expressão de FGFR-1 no epitélio apresentaram taxa de SDE em 5 anos de 22,9%, contra 75,6% dos pacientes com baixa expressão (HR: 2,594; IC95%: 1,390-4,841; p=0,003). O mesmo aconteceu com FGFR-1 no estroma, com taxa de SDE em 5 anos de 32,9%, contra 64,0% (HR: 3,378; IC95%: 1,816-6,286; p=0,001). A análise multivariada de Cox confirmou que a expressão de FGF-2 no estroma (HR: 2,197; IC95%: 1,128-4,282; p=0,02), FGFR-1 no epitélio (HR: 3,178; IC95%: 1,505-6,709; p=0,002) e FGFR-1 no estroma (HR: 3,041; IC95%: 1,454-6,356; p=0,003) estão fortemente associadas a um maior risco de morte relacionada ao CECL. Em conjunto, nossos achados demonstram que FGF-2 e FGFR-1 desempenham um papel importante na DEO e no CECL, estando associados à presença de metástase e à sobrevivência dos pacientesAbstract: Fibroblast growth factor 2 (FGF-2) and fibroblast growth factor receptor 1 (FGFR-1) expression is associated with tumour invasiveness, cell proliferation, angiogenesis and metastasis potential. This study aimed to investigate FGF-2 and FGFR-1 expression in oral epithelial dysplasia (OED) and tongue squamous cell carcinoma (TSCC). One hundred and sixty-seven cases were retrospectively selected, including 85 surgical specimens of patients with TSCC, from Onofre Lopes Hospital, Natal, Brazil, besides 46 TSCC and 36 OED incisional biopsies from the Laboratory of Oral Pathology, Piracicaba Dental School, University of Campinas (UNICAMP). Tissue sections were submitted to immunohistochemical reaction for FGF-2 and FGFR-1. Slides were scanned and the immunostaining was digitally quantified by the Aperio Positive Pixel Count v9 software. Five areas were selected from the stroma and the epithelium. Results were exported and the final score of each lesion was obtained by the sum of the percentage of weak, moderate and strong pixels, resulting in a value ranging from 100 to 300. Cases were classified as "weak expression" or "high expression" of the proteins, accordingly to the group median. FGF-2 and FGFR-1 were more expressed in high-grade OED than in low-grade OED, either in the stroma or in the epithelium (p<0.05). The 5-year disease-specific survival (DSS) rate was 47.3% of the patients with TSCC. FGF-2 high expression in the inflammatory and mesenchymal cells of the stroma was associated with vascular invasion and worse prognosis. Patients with high expression of FGF-2 in the stroma had a 5-year DSS of 36.7%, against 59.3% of patients with low expression (HR: 2.272; CI(95%): 1.213-4.254; p=0.008). FGFR-1 high expression in the stroma was correlated with lymph node metastasis and distant metastasis. Patients with high expression of FGFR-1 in the tumour had a 5-year DSS of 22.9%, against 75.6% of patients with low expression (HR: 2.594; CI(95%): 1.390-4.841; p=0.003). The same was observed with FGFR-1 in the stroma, with a 5-year DSS of 32.9%, against 64.0% (HR: 3.378; CI(95%): 1.816-6.286; p=0.001). The Cox multivariate analysis confirmed that the expression of FGF-2 in the stroma (HR: 2.197; CI(95%): 1.128-4.282; p=0.02), FGFR-1 in the tumour (HR: 3.178; CI(95%): 1.505-6.709; p=0.002) and FGFR-1 in the stroma (HR: 3.041; CI(95%): 1.454-6.356; p=0.003) are strongly associated with a higher risk of death related to TSCC. Taken together, our findings demonstrate that FGF-2 and FGFR-1 play an important role in OED and TSCC, and are associated with the presence of metastasis and patients disease-specific survivalMestradoEstomatopatologiaMestre em Estomatopatologia33003033009P4CAPE

Efeito do fumo na expressão proteica da citoqueratina 19 (k19) em células epiteliais da mucosa bucal

Objective: to compare the expression of cytokeratin 19 (K19) in epithelial cells of the oral mucosa of smokers, nonsmokers and former smokers. Methods: samples of epithelium of the oral mucosa of smokers (n=20), non-smokers (n=20) and former smokers (n=20) were collected by mouthwash and processed by immunohistochemistry. The number of stained cells was counted in five fields of slides prepared in duplicate. The number of stained cells was counted in five fields per slide by two observers (k = 1.00). Statistical analysis was performed using one-way ANOVA and Tukey tests at a significance level of 5%. Results: smokers had higher percentage of stained cells (74.5%) compared to non-smokers (35.0%) (p=0.003). Former smokers showed 55.3% of stained cells for K19, but there were no significant differences compared to non-smokers (p=0.18) or smokers (p=0.22). Conclusion: smoking is associated with changes in protein expression of cytokeratin 19 (K19), being the greater number of stained cells observed in smokers, indicating changes in epithelial maturation of these individuals.Objetivo: comparar a expressão da citoqueratina 19 (K19) em células epiteliais da mucosa oral entre fumantes, não fumantes e ex-fumantes. Metodologia: amostras de epitélio da mucosa bucal de indivíduos fumantes (n=20), não fumantes (n=20) e ex-fumantes (n=20) foram coletadas por bochecho e processadas por imuno-histoquímica. O número de células marcadas foi contado em cinco campos de cada lâmina por dois avaliadores (k=1,00). A análise estatística foi realizada através do teste ANOVA One-Way e Tukey ao nível de significância de 5%. Resultados: indivíduos fumantes apresentaram maior porcentagem de células marcadas (74,5%) em relação aos não fumantes (35,0%) (p=0,003). Os indivíduos ex-fumantes apresentaram 55,3% de células marcadas para a K19, porém não houve diferenças significativas em comparação aos não fumantes (p=0,18) ou aos fumantes (p=0,22). Conclusão: o hábito de fumar está associado a alteração no perfil de expressão proteica da Citoqueratina 19 (K19), sendo observado maior número de células marcadas em fumantes, indicando alterações na maturação epitelial desses indivíduos

Radiographic estimation of the growth rate of initially underdiagnosed ameloblastomas

To evaluate the specific growth rate (SGR) of ameloblastoma. Cases of ameloblastoma initially underdiagnosed (e.g. cases overlooked or diagnosed as reactive lesions) which had adequate radiographic documentation to evaluate their progression were retrospectively selected. Two panoramic radiographs were analyzed to determine the specific growth rate (SGR) of each tumor, defined as the logarithm of the ratio of final tumor area (when the diagnosis of ameloblastoma was made) to the initial tumor area (when the lesion was underdiagnosed), divided by the time interval between the radiographic images. The tumor area was measured using the software ImageJ. Twelve patients with mandibular ameloblastomas were selected, including 5 males and 7 females, with a mean age of 24.9 years (range: 14-61 years). In four cases, the lesion was associated with the crown of an impacted third molar. In three cases, it was initially diagnosed as a periapical lesion. Three cases were extrafollicular and were not noticed in the initial radiographs. Two cases were initially diagnosed as ameloblastoma, but the surgery was delayed for personal reasons. The mean interval of time between the two radiographic images was 4.3 years (range: 0.4-9 years). Based on our analysis, ameloblastoma grows in average 40.4% per year (range: 14.9-88.7%). Ameloblastoma is a progressively growing tumor, but its growth rate seems to be smaller than initially reported in the literature. Better understanding the radiographic progression of ameloblastoma might improve its early diagnosis, management, and prognosis

Polypodium leucotomos targets multiple aspects of oral carcinogenesis and it is a potential antitumor phytotherapy against tongue cancer growth

Peer reviewe

Heparanase 1 Upregulation Promotes Tumor Progression and Is a Predictor of Low Survival for Oral Cancer

Background: Oral cavity cancer is still an important public health problem throughout the world. Oral squamous cell carcinomas (OSCCs) can be quite aggressive and metastatic, with a low survival rate and poor prognosis. However, this is usually related to the clinical stage and histological grade, and molecular prognostic markers for clinical practice are yet to be defined. Heparanase (HPSE1) is an endoglycosidase associated with extracellular matrix remodeling, and although involved in several malignancies, the clinical implications of HPSE1 expression in OSCCs are still unknown.Methods: We sought to investigate HPSE1 expression in a series of primary OSCCs and further explore whether its overexpression plays a relevant role in OSCC tumorigenesis. mRNA and protein expression analyses were performed in OSCC tissue samples and cell lines. A loss-of-function strategy using shRNA and a gain-of-function strategy using an ORF vector targeting HPSE1 were employed to investigate the endogenous modulation of HPSE1 and its effects on proliferation, apoptosis, adhesion, epithelial-mesenchymal transition (EMT), angiogenesis, migration, and invasion of oral cancer in vitro.Results: We demonstrated that HPSE1 is frequently upregulated in OSCC samples and cell lines and is an unfavorable prognostic indicator of disease-specific survival when combined with advanced pT stages. Moreover, abrogation of HPSE1 in OSCC cells significantly promoted apoptosis and inhibited proliferation, migration, invasion, and epithelial-mesenchymal transition by significantly decreasing the expression of N-cadherin and vimentin. Furthermore, a conditioned medium of HPSE1-downregulated cells resulted in reduced vascular endothelial growth.Conclusion: Our results confirm the overexpression of HPSE1 in OSCCs, suggest that HPSE1 expression correlates with disease progression as it is associated with several important biological processes for oral tumorigenesis, and can be managed as a prognostic marker for patients with OSCC.Peer reviewe

Polypodium leucotomos targets multiple aspects of oral carcinogenesis and it is a potential antitumor phytotherapy against tongue cancer growth

Introduction: Oral cancer refers to malignant tumors, of which 90% are squamous cell carcinomas (OSCCs). These malignancies exhibit rapid progression, poor prognosis, and often mutilating therapeutical approaches. The determination of a prophylactic and/or therapeutic antitumor role of the polyphenolic extract Polypodium leucotomos(PL) would be relevant in developing new tools for prevention and treatment.Methods: We aimed to determine the antitumor effect of PL by treating OSCC cell lines with PL metabolites and evaluating its action during OSCC progression in vivo.Results: PL treatment successfully impaired cell cycling and proliferation, migration, and invasion, enhanced apoptosis, and modulated macrophage polarization associated with the tumoral immune-inflammatory response of tongue cancer cell lines (TSCC). PL treatment significantly decreased the expression of MMP1 (p &lt; 0.01) and MMP2 (p &lt; 0.001), and increased the expression of TIMP1 (p &lt; 0.001) and TIMP2 (p &lt; 0.0001) in these cells. The mesenchymal-epithelial transition phenotype was promoted in cells treated with PL, through upregulation of E-CAD (p &lt; 0.001) and reduction of N-CAD (p &lt; 0.05). PL restrained OSCC progression in vivo by inhibiting tumor volume growth and decreasing the number of severe dysplasia lesions and squamous cell carcinomas. Ki-67 was significantly higher expressed in tongue tissues of animals not treated with PL(p &lt; 0.05), and a notable reduction in Bcl2 (p &lt; 0.05) and Pcna (p &lt; 0.05) cell proliferation-associated genes was found in dysplastic lesions and TSCCs of PL-treated mice. Finally, N-cad(Cdh2), Vim, and Twist were significantly reduced in tongue tissues treated with PL.Conclusion: PL significantly decreased OSCC carcinogenic processes in vitro and inhibited tumor progression in vivo. PL also appears to contribute to the modulation of immune-inflammatory oral tumor-associated responses. Taken together, these results suggest that PL plays an important antitumor role in processes associated with oral carcinogenesis and may be a potential phytotherapeutic target for the prevention and/or adjuvant treatment of TSCC

A Rare Case of Mandibular Aspergillus Osteomyelitis in an Immunocompetent Patient

Aspergillosis is a fungal infection caused by Aspergillus species, which is contracted through spores that colonize the respiratory tract, causing rhinosinusitis and pulmonary infections. Oral aspergillosis is rare and, when present, may cause soft tissue and bone destruction, generally in immunodeficient patients. Mandibular Aspergillus osteomyelitis is even rarer, with few cases reported in the literature. A 57-year-old Caucasian woman was referred for the evaluation of painful recurrent swelling in the anterior mandibular alveolar ridge, with purulent drainage, previously treated with multiple surgical debridement procedures and antibiotics without success. The patient was otherwise systemically healthy. Surgical debridement was performed and histopathological examination showed osteomyelitis associated with Aspergillus species. Therapy with oral itraconazole (400 mg per day) was administered for 3 months, resulting in complete resolution. No recurrence was detected after 15 years of follow-up. The patient was rehabilitated with dental implants. In conclusion, non-bacterial microorganisms, such as Aspergillus, should be considered in cases of mandibular osteomyelitis that do not heal after surgical debridement and antibiotic therapy

Clinical predictors of malignancy in palatal salivary gland tumors

To establish a predictive clinical index of malignancy risk in palatal salivary gland tumors (PSGT). Materials and methods One hundred cases of PSGT were evaluated. Clinical data were retrieved from the patient's files. Representative clinical photographs of each tumor were evaluated to identify clinical features suggestive of a malignant tumor. Features significantly associated with malignancy were included in a binary logistic regression model. Results Malignant tumors were more common in the hard palate, in women and in older patients. Features associated with a malignant diagnosis included pain (p = .017), irregular surface (p = .004), bluish/purple coloration (p < .001), ulceration (p = .005), and telangiectasia (p = .015). After multivariate logistic regression, pain (OR: 4.017; 95% CI: 1.198-13.471; p = .024) and color alteration (OR: 7.243; 95% CI: 2.068-25.363; p = .002) were independently associated with malignancy. Including these factors in a predictive index, the proportion of malignant tumors in patients presenting none, one and two factors were 25% (95% CI: 0.13-0.40), 67% (95% CI: 0.48-0.83), and 85% (95% CI: 0.42-0.99), respectively. Conclusion Pain and color alteration might be independent predictors of malignancy in PSGT, which could support the decision to perform an incisional or excisional biopsy25819191924CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPE

Prognostic outcomes of oral squamous cell carcinoma derived from proliferative verrucous leukoplakia: A systematic review.

OBJECTIVE: This study aimed to evaluate prognostic outcomes of PVL-derived oral squamous cell carcinomas (P-OSCC) based on recurrence, new primary tumour, metastasis and survival information. STUDY DESIGN: Five databases and grey literature were searched electronically with the following main keywords (proliferative verrucous leukoplakia, squamous cell carcinoma and malignant transformation) to answer the following review question: &apos;Are survival outcomes for P-OSCC worse?&apos; based on the PECOS principle. The Joanna Briggs Institute Critical Appraisal tool was used to identify possible biases and assess the quality of each of the primary studies. RESULTS: A total of 21 articles met the inclusion criteria, and the results of this systematic review suggest that P-OSCC can recur and generate new primary tumours; however, metastases are rare. Thus, most patients remain alive for an average period of 5 years. CONCLUSION: Apparently, P-OSCC has better clinical prognostic characteristics than conventional OSCC. There is a lack of information on the main prognostic outcomes of P-OSCC; therefore, specific studies must be performed to achieve a better comparison between P-OSCC and conventional OSCC progression

Polypodium leucotomos targets multiple aspects of oral carcinogenesis and it is a potential antitumor phytotherapy against tongue cancer growth

Abstract Introduction: Oral cancer refers to malignant tumors, of which 90% are squamous cell carcinomas (OSCCs). These malignancies exhibit rapid progression, poor prognosis, and often mutilating therapeutical approaches. The determination of a prophylactic and/or therapeutic antitumor role of the polyphenolic extract Polypodium leucotomos(PL) would be relevant in developing new tools for prevention and treatment. Methods: We aimed to determine the antitumor effect of PL by treating OSCC cell lines with PL metabolites and evaluating its action during OSCC progression in vivo. Results: PL treatment successfully impaired cell cycling and proliferation, migration, and invasion, enhanced apoptosis, and modulated macrophage polarization associated with the tumoral immune-inflammatory response of tongue cancer cell lines (TSCC). PL treatment significantly decreased the expression of MMP1 (p &lt; 0.01) and MMP2 (p &lt; 0.001), and increased the expression of TIMP1 (p &lt; 0.001) and TIMP2 (p &lt; 0.0001) in these cells. The mesenchymal-epithelial transition phenotype was promoted in cells treated with PL, through upregulation of E-CAD (p &lt; 0.001) and reduction of N-CAD (p &lt; 0.05). PL restrained OSCC progression in vivo by inhibiting tumor volume growth and decreasing the number of severe dysplasia lesions and squamous cell carcinomas. Ki-67 was significantly higher expressed in tongue tissues of animals not treated with PL(p &lt; 0.05), and a notable reduction in Bcl2 (p &lt; 0.05) and Pcna (p &lt; 0.05) cell proliferation-associated genes was found in dysplastic lesions and TSCCs of PL-treated mice. Finally, N-cad(Cdh2), Vim, and Twist were significantly reduced in tongue tissues treated with PL. Conclusions: PL significantly decreased OSCC carcinogenic processes in vitro and inhibited tumor progression in vivo. PL also appears to contribute to the modulation of immune-inflammatory oral tumor-associated responses. Taken together, these results suggest that PL plays an important antitumor role in processes associated with oral carcinogenesis and may be a potential phytotherapeutic target for the prevention and/or adjuvant treatment of TSCCs