Paulo Antônio Silvestre de Faria

Expression of Bcl-2 family proteins and associated clinicopathologic factors predict survival outcome in patients with oral squamous cell carcinoma.

The aims of this study were to assess the expression levels of three proteins involved in apoptosis--Bcl-2, Bcl-X, and Bax--and evaluate their relationship with clinicopathologic features and survival in oral squamous cell carcinoma (OSCC). Immunohistochemistry was used to evaluate protein expression in 53 primary OSCCs treated by radical surgery with free margins at a single institution in 1999. Histologic specimens were graded and analyzed for perineural invasion, lymphocytic infiltrate, and pattern of invasion. Digital image analysis was performed to quantify immunostaining. Survival was analyzed using the Kaplan-Meier method and Coxs proportional hazard model. Cancer-specific 5-year survival (CSS) was 61% (56% overall survival (OS), and 51% disease-free interval (DFI)). Kaplan-Meier analysis identified pathologic stage (p=0.0007, log-rank test, OS), negative nodes status (pN) (p<0.0001, log-rank test, OS), presence of lymphocytic infiltrate (p=0.0084, log-rank test, OS), and high Bax expression (p=0.025, log-rank test, OS) to each be associated with both better OS and CSS. Tongue tumors (p=0.0179, log-rank test), worst pattern of invasion (p=0.0293, log-rank test), lack of lymphocytic infiltrate (p=0.0328, log-rank test), perineural invasion (p=0.0448, log-rank test), poorly differentiated tumors (p=0.0318, log-rank test), and low Bcl-X expression (p=0.044, log-rank test) were all associated with a low DFI. Cox regression found pN, lymphocytic infiltrate, and Bax expression to be independent prognostic factors for OS and CSS, whereas lymphocytic response and tongue tumors were predictors of DFI. Bcl-2 expression emerged as an independent marker of favorable CSS. Lymphocytic infiltrate was the most meaningful histopathologic parameter in survival analysis, whereas expression of Bcl-2 family members seems to be an important marker of a favorable prognosis in OSCC.

Oral Squamous Cell Carcinoma: Clinicopathological Features in Patients with and without Recurrence

Aim: To compare the clinicopathological profile of oral squamous cell carcinoma (OSCC) in groups with and without recurrence. Methods: Records of all patients who underwent surgery for primary OSCC at a single institution during 1999 were identified. Patient demographics, lesion site, clinical and pathologic stage, pathologic grading, pattern of invasion, lymphocytic infiltrate, perineural invasion, and treatment and survival data were collected. Descriptive statistics were calculated for each variable and survival was calculated using Kaplan-Meier and Cox models. Patients were divided into 2 groups: with (n = 25) and without (n = 28) recurrence. Results: Tongue (p = 0.02) and poorly differentiated (p = 0.04) tumors were associated with recurrence. Kaplan-Meier and Cox models revealed tobacco use and the absence of lymphocytic infiltrate to be associated with the poorest survival in recurrent OSCC. Conclusion: The tumor site, tobacco use, and pathological features were involved in the recurrence of OSCC and should be taken into account for OSCC treatment and follow-up.

HPV infection in Brazilian patients with esophageal squamous cell carcinoma: Interpopulational differences, lack of correlation with surrogate markers and clinicopathological parameters

The role of HPV in esophageal squamous cell carcinoma (ESCCs) is controversial. Therefore, we determined, through different methodologies, the prevalence of HPV in 264 ESCC samples from Brazil, and correlated it with the presence of surrogate markers and clinicopathological characteristics. HPV is present in 13% of ESCC, and with a 3-fold variation between high and medium incidence areas. Most HPV positive tumors were infected with HPV16, but this was not associated with p16 expression, TP53 mutation status, patient age, amount of tobacco or alcohol consumption, or overall survival. We conclude that HPV infection may not have a role in ESCC.

Alterations in epidermal growth factor receptors 1 and 2 in esophageal squamous cell carcinomas

A bstractBackgroundEsophageal squamous cell carcinoma (ESCC) shows a 5-year survival rate below 10%, demonstrating the urgency in improving its treatment. Alterations in epidermal growth factor receptors are closely related to malignancy transformation in a number of tumors and recent successful targeted therapies have been directed to these molecules. Therefore, in this study, we analyzed the expression of EGFR and HER2 and evaluated EGFR mutation profile as well as the presence of mutations in hotspots of KRAS and BRAF in ESCC patients.MethodsWe performed RT-qPCR, immunohistochemistry and Fluorescent in situ hybridization to determine EGFR and HER2 expression in ESCC patients, and direct sequencing and PCR-RFLP for mutations and polymorphism analysis.ResultsOur results showed an increased EGFR mRNA expression in tumors compared to surrounding tissue (p <0.05), with 11% of the cases presenting at least a four-fold difference between tumor and paired adjacent mucosa. EGFR protein overexpression was present only in 4% of the cases. The median expression of HER2 mRNA was not different between tumors and adjacent mucosa. Still, 7% of the tumors presented at least a 25-fold higher expression of this gene when compared to its paired counterpart. Immunohistochemical analysis revealed that 21% of the tumors were positive for HER2 (scores 2+ and 3+), although only 3+ tumors presented amplification of this gene. Mutation analysis for EGFR (exons 18-21), KRAS (codons 12 and 13) and BRAF (V600E) showed no mutations in any of the hotspots of these genes in almost 100 patients analyzed. EGFR presented synonymous polymorphisms at codon 836 (C>T) in 2.1% of the patients, and at codon 787 (G>A) in 79.2% of the cases. This last polymorphism was also evaluated in 304 healthy controls, which presented a similar frequency (73.7%) in comparison with ESCC patients. The absence of mutations of EGFR, KRAS and BRAF as well as the overexpression of EGFR and HER2 in less than 10% of the patients suggest that this signaling pathway is altered in only a small proportion of patients with ESCC.ConclusionHER receptors target therapies may have the potential to be effective in only a minor fraction of patients with ESCC.

SUBLINGUAL GLAND TUMORS: CLINICAL, PATHOLOGIC, AND THERAPEUTIC ANALYSIS OF 13 PATIENTS TREATED IN A SINGLE INSTITUTION

Sublingual gland tumors are rare, although frequently malignant. This study describes the clinicopathologic features and treatment results and reviews the literature.

Análise de sobrevida global em pacientes diagnosticados com carcinoma de células escamosas de boca no INCA no ano de 1999

O carcinoma de celulas escamosas de boca compreende cerca de 90 a 95% de todas as neoplasias malignas da boca e e um dos tipos de câncer mais frequentes no Brasil. O indice de sobrevida em 5 anos e baixo e permaneceu estavel nas ultimas decadas, apesar dos avancos nas terapias. O objetivo deste estudo foi analisar o perfil e a sobrevida global dos pacientes diagnosticados com carcinoma de celulas escamosas de boca no ano de 1999 no Instituto Nacional de Câncer. Dos 320 pacientes incluidos no estudo, 79,4% eram homens. A idade media foi de 56,7 anos, e 82,2% deles fumavam e/ou bebiam. A lingua, seguida do assoalho de boca foram os locais mais acometidos. A maioria (68,9%) dos pacientes foi diagnosticada em estadios tardios e submetida a radioterapia exclusiva (53,6%). A sobrevida media no periodo do estudo foi de 29,4 meses. Os pacientes dos estadios iniciais apresentaram maior sobrevida, assim como aqueles submetidos apenas a cirurgia como forma de tratamento e os que nao apresentaram linfonodos acometidos ao diagnostico. Tumores localizados em palato duro e mucosa jugal apresentaram pior prognostico. Foram fatores preditivos independentes de melhor sobrevida os tumores T1 ou T2 (p=0,001), sem acometimento de linfonodos (p=0,012) e nao localizados em mucosa jugal (p=0,021). O diagnostico do câncer oral ainda se faz em estadios tardios, o que influencia negativamente a sobrevida global dos pacientes. Maior enfase deve ser dada a capacitacao dos profissionais para o reconhecimento precoce do câncer e a conscientizacao da populacao de risco.

Vascular endothelial growth factor and thymidine phosphorylase expression in salivary gland tumors with distinct metastatic behavior

BACKGROUND Metastasis of salivary gland tumors has a negative impact on survival. Angiogenesis and its factors are potential markers for predicting metastasis in different malignant tumors, but this is not the case for salivary gland tumors. METHODS Salivary gland tumors of distinct biologic behavior were analyzed according to the semiquantitative immunoexpression of vascular endothelial growth factor (VEGF) and thymidine phosphorylase (TP). RESULTS Vascular endothelial growth factor expression was predominantly weak in benign tumors. Weak TP expression was observed in 100% cases of benign tumors and in 74.3% of primary malignant tumors. High VEGF and TP expression levels were significantly associated with primary malignant tumors but not with primary non-metastasizing and primary metastasizing malignant tumors or with subtypes of malignant tumors. CONCLUSIONS Vascular endothelial growth factor and TP expression levels discriminate benign and malignant tumors but cannot predict metastasis from non-metastasizing tumors.

Malignant peripheral nerve sheath tumors: clinicopathological aspects, expression of p53 and survival

Malignant peripheral nerve sheath tumors (MPNSTs) arerare and highly aggressive neoplasms, representing only 5%of soft tissue sarcomas (1,2). Approximately half of MPNSTcases occur in association with neurofibromatosis type 1(NF1) (3). MPNSTs may appear de novo or develop from themalignant transformation of a benign neural neoplasm,generally a plexiform neurofibroma (1). Solitary (unasso-ciated with NF1) and localized (or discrete; multiple in NF1)neurofibromas do not have malignant transformationpotential (1,3). NF1 loss of heterozygosity (LOH) has beendemonstrated in NF1-associated and sporadic MPNSTs.Although NF1 LOH is believed to be sufficient for neu-rofibroma development, MPNST pathogenesis has beensuggested to be a multistage process that includes othermolecular alterations (4,5). TP53 mutations have been foundin a subgroup of MPNSTs, indicating that a p53-mediatedpathway is involved in their development (5,6).Some clinicopathological features (e.g., the presence ofNF1,high histologicalgrade,necrosis,andrhabdomyoblasticdifferentiation) have been indicated to be important factorsforlower survival in MPNST cases in some studies butnotinothers (2,7–10). The clinical significance of p53 expression inMPNSTs is also a controversial issue. We aimed to study p53expression in MPNSTs and investigate its impact, as well astheimpactsoftheclinicopathologicalfeaturesofMPNSTs,onthe survival rates. We also compared p53 expression inMPNSTswith theirclinicopathological features and with p53expression in neurofibromas.

Quantitative evaluation of SPRR3 expression in esophageal squamous cell carcinoma by qPCR and its potential use as a biomarker

Esophageal squamous cell carcinoma (ESCC) is highly fatal due to late diagnosis and inefficient treatment. Early disease detection could improve diagnosis and patient survival. Esophageal squamous epithelial cells express SPRR3, a member of the small proline-rich protein family, which is downregulated in ESCC. Therefore, SPRR3 expression may be used as a biomarker to follow the transition from healthy mucosa to ESCC. Both SPRR3 mRNA splice variants, v1 and v2, were evaluated by real time PCR in tumor and histologically normal adjacent tissue biopsies from 84 ESCC patients and 18 healthy controls. SPRR3-v1 was most highly expressed in the esophageal mucosa of healthy subjects, with an increasingly lower expression in the adjacent mucosa of ESCC patients and in tumors, respectively. SPRR3-v2 expression was low in normal mucosa and in tumors but it was higher in the adjacent mucosa of ESCC patients. In addition, we found a significant correlation between a lower SPRR3-v1 and SPRR3-v2 expression and age and alcohol consumption, respectively. SPRR3 protein expression presented a good correlation with SPRR3 mRNA expression. Cut-off points to discriminate between healthy mucosa, tumor and adjacent mucosa were determined with receiver operating characteristic (ROC) curves. This analysis showed that SPRR3-v1 expression discriminates the esophageal mucosa of healthy subjects from the adjacent mucosa and the tumor of ESCC patients with high sensitivity and specificity. Our data shows that the quantitative analysis of SPRR3 mRNA is a robust and reliable method to monitor the malignant transformation of the healthy esophageal mucosa into ESCC.

Evaluation of Bcl-2, Bcl-x and cleaved caspase-3 in malignant peripheral nerve sheath tumors and neurofibromas.

AIMS To study the expression of Bcl-2, Bcl-x, as well the presence of cleaved caspase-3 in neurofibromas and malignant peripheral nerve sheath tumors. The expression of Bcl-2 and Bcl-x and the presence of cleaved caspase 3 were compared to clinicopathological features of malignant peripheral nerve sheath tumors and their impact on survival rates were also investigated. MATERIALS AND METHODS The evaluation of Bcl-2, Bcl-x and cleaved caspase-3 was performed by immunohistochemistry using tissue microarrays in 28 malignant peripheral nerve sheath tumors and 38 neurofibromas. Immunoquantification was performed by computerized digital image analysis. CONCLUSIONS Apoptosis is altered in neurofibromas and mainly in malignant peripheral nerve sheath tumors. High levels of cleaved caspase-3 are more common in tumors with more aggressive histological features and it is associated with lower disease free survival of patients with malignant peripheral nerve sheath tumors.