Cholestatic syndrome as initial manifestation of pancreatic metastasis of papillary thyroid carcinoma: case report and review.

Most papillary thyroid carcinomas (PTC) harbor excellent prognosis. Although rare, distant metastases normally occur in lungs and/or bones. Here we describe a rare case of pancreatic metastasis presenting with rapid onset cholestatic syndrome. A literature review was also performed. A 73-year-old man with a high risk PTC was submitted to total thyroidectomy (TT) followed by radioiodine therapy. After initial therapy, he persisted with progressive rising serum thyroglobulin levels but with no evidence of structural disease. Recently, the patient presented with a rapid onset and progressive cholestatic syndrome. A 4 cm lesion in pancreas was identified, with echoendoscopy fine-needle aspiration biopsy (FNAB) confirming a pancreatic metastasis from PTC. The patient was submitted to a successful pancreaticoduodenectomy. Pancreatic metastases of PTC are rare and few long-term follow-up data are available to guide management. Fourteen cases were former reported, mean age was 65.7 years-old with mean time between PTC and pancreatic metastasis diagnosis of 7.9 years. Nine of them had another distant metastasis, nine were diagnosed by FNAB and just two received sorafenib.

Cholestatic syndrome as initial manifestation of pancreatic metastasis of papillary thyroid carcinoma: case report and review

SUMMARY Most papillary thyroid carcinomas (PTC) harbor excellent prognosis. Although rare, distant metastases normally occur in lungs and/or bones. Here we describe a rare case of pancreatic metastasis presenting with rapid onset cholestatic syndrome. A literature review was also performed. A 73-year-old man with a high risk PTC was submitted to total thyroidectomy (TT) followed by radioiodine therapy. After initial therapy, he persisted with progressive rising serum thyroglobulin levels but with no evidence of structural disease. Recently, the patient presented with a rapid onset and progressive cholestatic syndrome. A 4 cm lesion in pancreas was identified, with echoendoscopy fine-needle aspiration biopsy (FNAB) confirming a pancreatic metastasis from PTC. The patient was submitted to a successful pancreaticoduodenectomy. Pancreatic metastases of PTC are rare and few long-term follow-up data are available to guide management. Fourteen cases were former reported, mean age was 65.7 years-old with mean time between PTC and pancreatic metastasis diagnosis of 7.9 years. Nine of them had another distant metastasis, nine were diagnosed by FNAB and just two received sorafenib.

How pathological criteria can impact prognosis of tongue and floor of the mouth squamous cell carcinoma.

other: Pathological parameters have been indicated as tumor prognostic factors in oral carcinoma. OBJECTIVE: The objective of this study was to investigate the impact of pathological parameters on prognosis of patients affected only by tongue and/or floor of the mouth squamous cell carcinoma (SCC). METHODOLOGY: In total, 380 patients treated in the Brazilian National Cancer Institute (INCA) from 1999 to 2006 were included. These patients underwent radical resection followed by neck dissection. The clinical and pathological characteristics were recorded. The Kaplan-Meier method and Cox proportional hazards model were used in survival analysis. Overall survival (OS), cancer-specific survival (CSS) and disease-free interval (DFI) were estimated. Cox residuals were evaluated using the R software version 3.5.2. Worst OS, CSS and DFI were observed in patients with tumors in advanced pathological stages (p<0.001), with the presence of perineural invasion (p<0.001) and vascular invasion (p=0.005). RESULTS: Advanced pathological stage and the presence of a poorly differentiated tumor were independent prognostic factors for OS and CSS. However, advanced pathological stage and perineural invasion were independent predictors of a shorter OS, DFI and CSS. CONCLUSION: Pathological stage and perineural invasion were the most significant pathological variables in survival analysis in tongue and/or floor of the mouth SCC.

How pathological criteria can impact prognosis of tongue and floor of the mouth squamous cell carcinoma

Abstract Pathological parameters have been indicated as tumor prognostic factors in oral carcinoma. Objective: The objective of this study was to investigate the impact of pathological parameters on prognosis of patients affected only by tongue and/or floor of the mouth squamous cell carcinoma (SCC). Methodology: In total, 380 patients treated in the Brazilian National Cancer Institute (INCA) from 1999 to 2006 were included. These patients underwent radical resection followed by neck dissection. The clinical and pathological characteristics were recorded. The Kaplan-Meier method and Cox proportional hazards model were used in survival analysis. Overall survival (OS), cancer-specific survival (CSS) and disease-free interval (DFI) were estimated. Cox residuals were evaluated using the R software version 3.5.2. Worst OS, CSS and DFI were observed in patients with tumors in advanced pathological stages (p<0.001), with the presence of perineural invasion (p<0.001) and vascular invasion (p=0.005). Results: Advanced pathological stage and the presence of a poorly differentiated tumor were independent prognostic factors for OS and CSS. However, advanced pathological stage and perineural invasion were independent predictors of a shorter OS, DFI and CSS. Conclusion: Pathological stage and perineural invasion were the most significant pathological variables in survival analysis in tongue and/or floor of the mouth SCC.

Twist1 Correlates With Epithelial-Mesenchymal Transition Markers Fibronectin and Vimentin in Adrenocortical Tumors.

BACKGROUND/AIM: Although the knowledge regarding adrenocortical carcinomas (ACC) tumorigenesis has significantly improved during the last decade, it still remains to be completely determined. Epithelial-mesenchymal transition (EMT) is a well described transcription factor induced process, postulated as an essential step toward cancer progression and metastasis development. In this context, Twist1 has been described as the EMT master-regulator. The aim of this study was to assess the association among Twist1, fibronectin, vimentin and E-cadherin gene expression in adrenocortical tumor samples. MATERIALS AND METHODS: Twist1, fibronectin, vimentin and E-cadherin gene expression in 18 adrenal adenomas, 18 ACC, and 24 childhood onset adrenocortical tumors were assessed in formalin-fixed paraffin-embedded tissues. The fold expression was calculated according to the 2ΔCt method. RESULTS: A significant correlation between mRNA levels of Twist1, fibronectin and vimentin was evident. Although their expression was inversely proportional, no association was observed between Twist1 and E-cadherin expression. CONCLUSION: The expression of Twist1, the major regulator of EMT, is directly correlated to the expression of mesenchymal markers fibronectin and vimentin in ACC samples.

Hamartoma condromatoso de parede torácica: relato de caso / Chondromatous chest wall hamartoma: case report

"O hamartoma condromatoso (HC) é um tipo raro de tumor benigno de cartilagem que se origina nos primeiros anos de vida. Na maioria das vezes seu sítio primário é o gradil torácico e pode ou não ter sintomatologia. Suas características histológicas mostram proliferação do tecido conjuntivo cartilaginoso associado a cistos de células ósseas. É de difícil diagnóstico devido sua apresentação histológica. Neste trabalho é relatado um caso de HC e sua evolução"

"Chondromatous hamartoma (HC) is a rare type of benign cartilage tumor that originates in the first years of life. Most of the time its primary site is the thoracic bar and may or may not have symptoms. Its histological characteristics show proliferation of cartilaginous connective tissue associated with bone cell cysts. It is difficult to diagnose due to its histological presentation. This work reports a case of HC and its evolution"

Alterações Citogenético-Moleculares no Gene FOXO1 em uma Criança com Rabdomiossarcoma Alveolar: Relato de Caso / Citogenetic-Molecular Alterations in FOXO1 Gene in a Child with Alveolar Rhabdomyosarcoma: Case Report / Cambios Citogenetico-moleculares en el gene fOxO1 en un Niño con Rabdomiossarcoma Alveolar: Relato de Caso

Introdução: o rabdomiossarcoma (rMs) é o tumor de tecidos moles mais comum da infância. Pode ser classificado em dois subtipos principais: o rabdomiossarcoma alveolar (rMsa) e o embrionário (rMse). no rMsa, o prognóstico é desfavorável quando comparado ao rMse, necessitando de tratamento intensificado; dessa forma, a distinção entre ambos os subtipos é fundamental. citogeneticamente, o rMsa apresenta translocações cromossômicas envolvendo o gene FOXO1 em 80% dos casos. a metodologia de hibridização in situ por fluorescência (FisH) tem sido muito utilizada para caracterizar o rMsa. Relato do caso: Paciente do sexo feminino, com 7 anos de idade, apresentou ao diagnóstico rMsa parameníngeo, sem metástase ao diagnóstico. a análise por meio de FisH mostrou a translocação envolvendo o gene FOXO1 e uma cópia extra desse gene. a paciente foi incluída no protocolo de tratamento do epssG, classificada como grupo de alto risco e recebeu quimioterapia e radioterapia. no final do tratamento, foi observada resposta parcial e iniciada quimioterapia de segunda linha. não houve resposta clinicorradiológica e a paciente evoluiu com progressão de doença local refratária ao tratamento e óbito após um ano do diagnóstico. Conclusão: de acordo com o nosso conhecimento, é a primeira descrição de um caso de rMsa apresentando a translocação do gene FOXO1 e uma cópia extra desse gene em clones separados. são necessários ainda novos estudos, a fim de compreender melhor o significado prognóstico da presença dessas alterações.

Introduction:rhabdomyosarcoma (rMs) is the most common soft tissue tumor of childhood. it can be classified into two main subtypes: alveolar rhabdomyosarcoma (arMs) and embryonal (erMs). in arMs the prognosis is unfavorable when compared to erMs, requiring intensified treatment, thus the distinction between both subtypes is fundamental. cytogenetically, arMs present chromosomal translocations involving the FOXO1 gene in 80% of the cases. The fluorescence in situ hybridization methodology (FisH) has been widely used to characterize arMs subtype. Case Report: a 7-year-old female patient presented with parameningeal arMs, non-metastatic at diagnosis. FisH analysis showed translocation involving the FOXO1 gene and an extra copy of this gene. The patient was enrolled in the epssG treatment protocol, classified as a high-risk group and received chemotherapy and radiotherapy. at the end of treatment a partial response was observed, and second line chemotherapy was started. There was no clinical-radiological response and the patient progressed with local disease, refractory to rescue treatment and died of disease one year after diagnosis. Conclusion:to our knowledge, this is the first case of arMs presenting FOXO1 gene translocation and an extra copy of this gene in separate clones. More studies are necessary to understand the prognostic significance of these alterations

Introducción: el rabdomiosarcoma (rMs) es el tumor de tejidos blandos más común de la infancia. el rMs puede clasificarse en dos subtipos principales, el rabdomiosarcoma alveolar (rMsa) y el embrionario (rMse). el rMsa presenta un pronóstico desfavorable si se compara al rMse, habiendo así necesidad de intensificación del tratamiento. de esta forma, la distinción entre rMsa y rMse es fundamental. citogéticamente, el rMsa presenta en cerca del 80% de los casos de translocación cromosómica que involucra el gen FOXO1. la metodología de Hibridación fluorescente in situ (FisH) ha sido muy utilizada para caracterizar el rMsa. Caso de estudio: Paciente del sexo femenino, de 7 años de edad presentada con un diagnóstico de rMsa parameningeo, sin metástasis. el análisis a través del FisH mostró la translocación envolviendo el gen FOXO1 y una copia extra de este gen. la paciente fue incluida en el protocolo de tratamiento del epssG, clasificado como grupo de alto riesgo y recibió quimioterapia y radioterapia. al final del tratamiento fue observada una respuesta parcial y se inició la quimioterapia de segunda línea. no hubo respuesta clínico-radiológica y la paciente evolucionó con progresión de enfermedad local, refractaria y óbito después de 1 año del diagnóstico. Conclusión: de acuerdo con nuestro conocimiento, este es el primer caso de un niño con rMsa presentando la translocación del gen FOXO1 y una copia extra de este gen en clones separados. se necesitan nuevos estudios para comprender mejor el significado pronóstico de la presencia de estos cambios.

Tumor de Wilms Bilateral Sincrônico: Avaliação Cirúrgica e Sobrevida / Synchronous Bilateral Wilms Tumor: Surgical Evaluation and Survival / Tumor de Wilms bilateral Sincrónico: Evaluación quirúrgica y Sobrevida

Introdução: os tumores renais representam cerca de 7% de todas as neoplasias malignas da infância. o tumor de Wilms bilateral sincrônico corresponde a 5-7% de todos os nefroblastomas. o tratamento consiste em quimioterapia pré-operatória, seguida de cirurgia conservadora, quimioterapia pós-operatória e, quando indicada, radioterapia. Objetivo: analisar o tipo de cirurgia e a sobrevida global dos pacientes com tumor de Wilms bilateral sincrônico. Método: este estudo de coorte retrospectivo incluiu 18 pacientes pediátricos do Hospital do câncer i, do instituto nacional de câncer José alencar Gomes da silva, no rio de Janeiro, de janeiro de 2000 a dezembro de 2017. curvas de sobrevida foram calculadas pelo método Kaplan-Meier. Resultados: a idade mediana ao diagnóstico foi de 19 meses, sendo dez casos do sexo feminino. em 62,5% dos rins operados, foi possível realizar cirurgia conservadora e, em 41,2% (7/17) dos pacientes, em ambos os rins. a sobrevida global em cinco anos, segundo o tipo cirurgia, foi de 87,5% para cirurgia conservadora e de 62,8% para nefrectomia total (p=0,0001). a sobrevida global em cinco anos para a coorte inteira foi de 70,8%. Conclusão: a cirurgia preservadora de tecido renal em crianças com o tumor de Wilms bilateral sincrônico é viável e pode ser realizada com segurança em centros de referência.

Introduction: Kidney tumors account for about 7% of all childhood malignancies. synchronous bilateral Wilms tumor corresponds to 5-7% of all nephroblastomas. The treatment consists of preoperative chemotherapy, followed by conservative surgery, postoperative chemotherapy and, when indicated, radiotherapy. Objective: to analyze the type of surgery and the overall survival of patients with synchronous bilateral Wilms tumor. Method: This retrospective cohort study included 18 pediatric patients from the Hospital do cancer i, from the national cancer institute José alencar Gomes da silva, in rio de Janeiro, from January 2000 to december 2017. survival curves were calculated using the Kaplan-Meier method. Results:The median age at diagnosis was 19 months, and 10 patients were female. it was possible to perform conservative surgery in 62.5% of the kidneys, and in 41.2% (7/17) of patients in both kidneys. The 5-year overall survival according to the type of surgery was 87.5% for conservative surgery and 62.8% for radical nephrectomy (p=0.0001). The 5-year overall survival of the entire cohort was 70.8%. Conclusion: conservative surgery in pediatric synchronous bilateral Wilms tumor is feasible and can be performed safely in reference centers.

Introducción:los tumores renales representan alrededor del 7% de todas las neoplasias malignas de la infancia. el tumor de Wilms bilateral sincrónico corresponde al 5-7% de todos los nefroblastomas. el tratamiento consiste en quimioterapia preoperatoria, seguida de cirugía conservadora, quimioterapia postoperatoria y, cuando indicada, la radioterapia. Objetivo: analizar el tipo de cirugía y la supervivencia global de los pacientes con un tumor de Wilms bilateral sincrónico. Método: este estudio de cohorte retrospectivo incluyó 18 pacientes pediátricos del Hospital del cáncer i, del instituto nacional de cáncer José alencar Gomes da silva, nel río de Janeiro, de enero de 2000 a diciembre de 2017. las curvas de sobrevida fueron calculadas por el método Kaplan-Meier. Resultados: la edad media al diagnóstico fue de 19 meses, siendo diez casos del sexo femenino. en el 62,5% de los riñones operados fue posible realizar cirugía conservadora, siendo en el 41,2% (7/17) de los pacientes en los dos riñones. la supervivencia global en cinco años según el tipo de cirugía fue de 87,5% para cirugía conservadora y de 62,8% nefrectomía total (p=0,0001). la supervivencia global en cinco años para la cohorte entera fue del 70,8%. Conclusión: la cirugía conservadora en niños con el tumor de Wilms bilateral sincrónico es viable y puede ser realizada con seguridad en centros de referencia

Association between long interspersed nuclear element-1 methylation levels and relapse in Wilms tumors.

BACKGROUND: Wilms tumor (WT) is a curable pediatric renal malignancy, but there is a need for new molecular biomarkers to improve relapse risk-directed therapy. Somatic alterations occur at relatively low frequencies whereas epigenetic changes at 11p15 are the most common aberration. We analyzed long interspersed element-1 (LINE-1) methylation levels in the blastemal component of WT and normal kidney samples to explore their prognostic significance. RESULTS: WT samples presented a hypomethylated pattern at all five CpG sites compared to matched normal kidney samples; therefore, the averaged methylation levels of the five CpG sites were used for further analyses. WT presented a hypomethylation profile (median 65.0%, 47.4-73.2%) compared to normal kidney samples (median 71.8%, 51.5-77.5%; p < 0.0001). No significant associations were found between LINE-1 methylation levels and clinical-pathological characteristics. We observed that LINE-1 methylation levels were lower in tumor samples from patients with relapse (median methylation 60.5%) compared to patients without relapse (median methylation 66.5%; p = 0.0005), and a receiving operating characteristic curve analysis was applied to verify the ability of LINE-1 methylation levels to discriminate WT samples from these patients. Using a cut-off value of 62.71% for LINE-1 methylation levels, the area under the curve was 0.808, with a sensitivity of 76.5% and a specificity of 83.3%. Having identified differences in LINE-1 methylation between WT samples from patients with and without relapse in this cohort, we evaluated other prognostic factors using a logistic regression model. This analysis showed that in risk stratification, LINE-1 methylation level was an independent variable for relapse risk: the lower the methylation levels, the higher the risk of relapse. The logistic regression model indicated a relapse risk increase of 30% per decreased unit of methylation (odds ratio 1.30; 95% confidence interval 1.07-1.57). CONCLUSION: Our results reinforce previous data showing a global hypomethylation profile in WT. LINE-1 methylation levels can be suggested as a marker of relapse after chemotherapy treatment in addition to risk classification, helping to guide new treatment approaches.

Is there a role for epithelial-mesenchymal transition in adrenocortical tumors?

PURPOSE: Epithelial-mesenchymal transition (EMT) is a biological dynamic process by which epithelial cells lose their epithelial phenotype and acquire mesenchymal invasive and migratory characteristics. This has been postulated as an essential step during cancer progression and metastasis. Although this is well described in other tumors, the role of EMT in adrenocortical tumors (ACT) has yet to be addressed. METHODS: The aim of this study was to evaluate the expression of EMT markers e-cadherin, vimentin, and fibronectin, along with EMT-transcription factors (EMT-TFs), TWIST1, SIP1, and SNAIL in 24 adrenocortical carcinoma (ACC), 19 adrenocortical adenomas (ACA), 27 childhood-onset adrenocortical tumors (CAT), and 12 normal adrenal glands. The association of EMT and EMT-TFs with clinical outcomes and pathology features were also evaluated. RESULTS: Cytoplasmic vimentin expression was increased among CAT samples when compared to ACC, ACA, and normal adrenal samples (p < 0.001). There was no difference in e-cadherin and fibronectin expression observed between groups. Nuclear and cytoplasmic expression of TWIST1 and SIP1 was stronger in CAT and ACC vs. ACA and normal tissue samples (all, p < 0.05). ACT, regardless of classification, exhibited increased SNAIL expression when compared to normal tissue (p < 0.05). A significant correlation was observed between vimentin and TWIST1 (r s = 0.44, p < 0.001); SIP1 (r s = 0.51, p < 0.001); and SNAIL (r s = 0.23, p < 0.05). TWIST1 and SIP1 expressions demonstrated a significant correlation (r s = 0.56, p < 0.001). High SIP1 expression was associated with a lower survival rate among ACC cases (p < 0.05). CONCLUSIONS: Vimentin, TWIST1, and SIP1 expressions are increased in aggressive ACT. Therefore, EMT may play a relevant role in adrenal tumorigenesis.

A Novel TP53 Mutation Associated with TWIST1 and SIP1 Expression in an Aggressive Adrenocortical Carcinoma.

Adrenocortical carcinomas (ACC) are very rare tumors related to TP53 mutations mostly in childhood onset cases. Epithelial-mesenchymal transition (EMT) transcription factors TWIST1 and Smad interacting protein 1 (SIP1) are related to poorer outcomes in other malignancies, but their role in ACC is unknown. We describe a case of an advanced metastatic ACC (Weiss-score of 9) in a patient at age 76. After primary tumor resection, mitotane therapy was started as palliation to low-volume liver metastasis. After a 2-year period of stable disease, the patient died due to brain metastasis. Somatic gene sequencing revealed a novel TP53 mutation in DNA extracted from paraffin-embedded tissue, a deletion of 8bp in exon 8 (c.811_818del8; GAGGTGCG/-) in homo or hemizygosis causing a subsequent frameshift and premature stop codon at position 302. Immunohistochemistry of P53 and p-Ser-15 P53 showed absent tumoral staining. In addition, immunohistochemical analysis showed an increased expression of the mesenchymal markers vimentin and fibronectin. At last, EMT transcription factors TWIST1 and SIP1 were also overexpressed in tumoral cells. This case report describes an aggressive ACC with not only a novel somatic mutation, but also a novel International Agency for Research on Cancer database 8 base-pair deletion in TP53 exon 8. In addition, the expression of EMT inducers TWIST1 and SIP1 have been reported for the first time in an ACC case. Further investigation is needed to clarify the biologic significance of this new TP53 mutation and its role in the EMT process.

A proteomic approach to compare saliva from individuals with and without oral leukoplakia.

BACKGROUND: Oral leukoplakia is the most common potentially malignant disorder in the oral cavity and can precede carcinoma. This study aimed to identify possible oral leukoplakia salivary biomarkers. METHODS: Unstimulated saliva was collected from participants and protein concentration was determined. Proteins were then precipitated with cold acetone and separated using 2DE over a pH range of 3-10. Spot demarcation and matching were performed and protein identification was done through MS analysis. Oral leukoplakia tissues were submitted to immunohistochemistry analysis for keratin 10 (CK10). A complementary analysis of oral leukoplakias that were not included previously was performed in addition. RESULTS: 226±10 spots were identified in oral leukoplakia 2DE gels, and 262±12 spots were identified in volunteers. Twenty-two spots were highly abundant in oral leukoplakias or not detected in the control group, such as apolipoprotein A1, alpha amylase, cystatins, keratin 10, and lysozyme precursor. All were identified. All oral leukoplakia cases were immunopositive for CK10, mainly in the superficial epithelial layers. CONCLUSIONS: The 2DE salivary protein profiles of individuals with and without oral leukoplakia were observably different. CK10 appears to be an interesting protein and should be further studied in oral carcinogenesis. SIGNIFICANCE: MS-based proteomics enables large-scale analysis of proteins. Proteomics can provide detailed descriptions of proteomes of cells and tissues, including body fluids, and appears as a powerful tool to study human disorders. Saliva is readily accessible through non invasive collection and can mirror diverse disease states. Saliva from both diseased and healthy subjects can be analyzed through 2DE and differences between groups could be found. Routine immunohistochemistry analysis confirmed one of these findings, with CK10 being positive tissues from individuals with oral leukoplakia. Therefore, the present study allows insights into development of an important potential oral cancer precursor, named oral leukoplakia. However, the results can be extrapolated and tested in other precancer states, such as proliferative verrucous leukoplakia, patients at risk of oral cancer due to lifestyle behavior and/or cancer history in the family or even those who are under surveillance after a treated primary oral cancer.

Recurrence and mortality prognostic factors in childhood adrenocortical tumors: Analysis from the Brazilian National Institute of Cancer experience.

Prognostic markers that can help identifying precocious risk of unfavorable outcomes in patients with childhood adrenocortical tumors (ACTs) are still unclear. This observational and retrospective study aimed to identify clinical and pathology prognostic factors of recurrence and death in a tertiary cancer center population. Clinical, pathology, demographic, staging, and therapy data from patients with childhood ACT (median age: 3.6 years) treated at the Brazilian National Institute of Cancer between 1997 and 2015 were assessed. Univariate and bivariate analyses were used to study the association of clinical and pathology characteristics with recurrence and mortality. Recurrence and disease-related mortality were the main outcomes. Twenty-seven patients were included. Complete tumor resection was performed in 21 cases. The median tumor size was 8.2 cm. Mitotane was the most common adjuvant/palliative therapy (n = 13). Recurrence occurred in 6 patients, after a median time of 7.2 months, and was more common among those with larger tumors (P =.008), higher Weiss score (P =.001), and microscopic tumoral necrosis (P =.002). Ten patients died from the disease. Older age (P =.04), larger tumor size (P =.002), metastatic disease (P =.003), previous recurrence (P =.003), incomplete resection (P =.002), intraoperative tumor spillage (P =.005), higher Weiss score (P =.03), microscopic necrosis (P =.005), and capsular invasion (P =.02) were all associated with increased death risk. Even though complete tumor resection was performed in most cases, a considerable number of cases of childhood ACT resulted in recurrence and death. Early identification of unfavorable outcomes is essential to determine ideal therapy and appropriate surveillance.

Array CGH Analysis of Paired Blood and Tumor Samples from Patients with Sporadic Wilms Tumor.

Wilms tumor (WT), the most common cancer of the kidney in infants and children, has a complex etiology that is still poorly understood. Identification of genomic copy number variants (CNV) in tumor genomes provides a better understanding of cancer development which may be useful for diagnosis and therapeutic targets. In paired blood and tumor DNA samples from 14 patients with sporadic WT, analyzed by aCGH, 22% of chromosome abnormalities were novel. All constitutional alterations identified in blood were segmental (in 28.6% of patients) and were also present in the paired tumor samples. Two segmental gains (2p21 and 20q13.3) and one loss (19q13.31) present in blood had not been previously described in WT. We also describe, for the first time, a small, constitutive partial gain of 3p22.1 comprising 2 exons of CTNNB1, a gene associated to WT. Among somatic alterations, novel structural chromosomal abnormalities were found, like gain of 19p13.3 and 20p12.3, and losses of 2p16.1-p15, 4q32.5-q35.1, 4q35.2-q28.1 and 19p13.3. Candidate genes included in these regions might be constitutively (SIX3, SALL4) or somatically (NEK1, PIAS4, BMP2) operational in the development and progression of WT. To our knowledge this is the first report of CNV in paired blood and tumor samples in sporadic WT.

Padronização da análise de mutações no gene TP53 em carcinomas de células escamosas de boca em material parafinado / Standardization of TP53 gene mutations analysis on oral squamous cell carcinoma from paraffin-embedded tissues

Introduction: The tumor protein p53 gene (TP53) is a constant target of investigation in cancer pathogenesis. Analysis by immunohistochemistry provides limited data about p53 in oral carcinogenesis, and TP53 sequencing can contribute to this analysis. However, obtaining high-quality and contamination-free deoxyribonucleic acid (DNA) for a proper amplification can be a difficult task when using paraffin-embedded tissues. Objective: Standardize DNA extraction, polymerase chain reaction (PCR) amplification and DNA sequencing techniques for TP53 mutation analysis. Material and methods: Thirty-nine cases of oral squamous cell carcinoma (OSCC) were selected from the Pathology Division of Instituto Nacional de Câncer (Inca). The DNA extraction method used was the QIAamp® DNA minikit® system. After DNA quantification by spectrophotometry, 250 ng of genetic material obtained from TP53 gene were amplified by PCR for exon 2 and by nested PCR for exon 6. Out of the total sample, 11 cases were selected for exon 2 sequencing. Results: The DNA samples presented mean concentration of 119.74 ± 88.86 ng/µl (28.9-556.4) and purity of 1.69 ± 0.18 (1-1.9). Thirty-three (84.6%) samples were amplified for exon 2, and all samples for exon 6 (39/100%). Readable sequencing data were obtained in 10 (90.9%) cases. Conclusion: Optimization of conditions for TP53 sequencing was obtained, and this will facilitate the analysis of mutations in paraffin-embedded tissues, allowing molecular retrospective studies...

Introdução: O gene TP53 (proteína tumoral p53) é alvo constante de investigação na patogênese do câncer. A imuno-histoquímica fornece dados limitados na análise de p53 no processo da carcinogênese bucal e o sequenciamento de TP53 pode contribuir nessa investigação. Contudo, a obtenção de ácido desoxirribonucleico (DNA) com qualidade para amplificação e livre de contaminação pode constituir uma tarefa difícil na utilização de material parafinado. Objetivo: Padronizar as técnicas de extração de DNA, amplificação por reação em cadeia da polimerase (PCR) e sequenciamento para a análise de mutações em TP53. Material e métodos: Foram selecionados 39 casos de carcinomas de células escamosas bucal da Divisão de Patologia do Instituto Nacional de Câncer (Inca). O DNA foi extraído utilizando o sistema comercial QIAamp® DNA minikit®. Após quantificação do DNA por espectrofotometria, 250 ng de amostra foram amplificados pela técnica de PCR para o éxon 2 e por nested PCR para o éxon 6 do gene TP53. Da amostra total, 11 casos foram selecionados para a padronização da reação de sequenciamento do éxon 2. Resultados: As amostras de DNA apresentaram concentração média de 119,74 ng/µl ± 88,86 (28,9-556,4 ng/µl) e pureza de 1,69 ± 0,18 (1-1,9). Do total das amostras analisadas, 33 (84,6%) foram amplificadas para o éxon 2, e todas (39/100%), para o éxon 6. No sequenciamento do éxon 2 obtiveram-se sequências passíveis de leitura em 10 (90,9%) casos. Conclusão: A otimização das condições para o sequenciamento de TP53 foi obtida, o que facilitará a análise de mutações em tecidos parafinados, permitindo...

The significance of p53 immunoexpression with different clones (DO-7 and PAb-240) in oral squamous cell carcinoma.

BACKGROUND/AIM: The TP53 gene is a tumor suppressor gene. Its product is a nuclear protein that regulates cell cycle arrest, apoptosis and DNA repair. Anti-p53 clones DO-7 and PAb-240 recognize the amino acid sequences 21-25 and 213-217, respectively. This study aimed to evaluate the expression of these clones and their relationship with clinicopathological features and survival analysis in oral squamous cell carcinomas (OSCC). METHODS: Information on 53 primary OSCC was collected at the Brazilian National Cancer Institute. An immunohistochemical method was applied to evaluate p53 expression (DO-7 and PAb-240). Their expression was analyzed quantitatively and correlated with clinicopathological features. Kaplan-Meier survival curves and log rank test were used. RESULTS: Immunopositivity for DO-7 was present in 64% of the cases, while 58% were positive for PAb-240. There was no correlation between immunoexpression of both antibodies and clinicopathological features or survival analysis. DO-7 expression was significantly higher (p = 0.001) than that of PAb-240. CONCLUSIONS: There were quantitative differences between the expression of the antibodies studied, which may reflect a different specificity of each one. To confirm immunohistochemical results and estimate the true prognostic role of TP53 in OSCC, it is important to perform mutation analysis.

Contribution of multiparameter flow cytometry immunophenotyping to the diagnostic screening and classification of pediatric cancer.

Pediatric cancer is a relatively rare and heterogeneous group of hematological and non-hematological malignancies which require multiple procedures for its diagnostic screening and classification. Until now, flow cytometry (FC) has not been systematically applied to the diagnostic work-up of such malignancies, particularly for solid tumors. Here we evaluated a FC panel of markers for the diagnostic screening of pediatric cancer and further classification of pediatric solid tumors. The proposed strategy aims at the differential diagnosis between tumoral vs. reactive samples, and hematological vs. non-hematological malignancies, and the subclassification of solid tumors. In total, 52 samples from 40 patients suspicious of containing tumor cells were analyzed by FC in parallel to conventional diagnostic procedures. The overall concordance rate between both approaches was of 96% (50/52 diagnostic samples), with 100% agreement for all reactive/inflammatory and non-infiltrated samples as well as for those corresponding to solid tumors (n = 35), with only two false negative cases diagnosed with Hodgkin lymphoma and anaplastic lymphoma, respectively. Moreover, clear discrimination between samples infiltrated by hematopoietic vs. non-hematopoietic tumor cells was systematically achieved. Distinct subtypes of solid tumors showed different protein expression profiles, allowing for the differential diagnosis of neuroblastoma (CD56(hi)/GD2(+)/CD81(hi)), primitive neuroectodermal tumors (CD271(hi)/CD99(+)), Wilms tumors (>1 cell population), rhabdomyosarcoma (nuMYOD1(+)/numyogenin(+)), carcinomas (CD45(-)/EpCAM(+)), germ cell tumors (CD56(+)/CD45(-)/NG2(+)/CD10(+)) and eventually also hemangiopericytomas (CD45(-)/CD34(+)). In summary, our results show that multiparameter FC provides fast and useful complementary data to routine histopathology for the diagnostic screening and classification of pediatric cancer.

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

BACKGROUND: Esophageal 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. METHODS: We 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. RESULTS: Our 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. CONCLUSION: HER receptors target therapies may have the potential to be effective in only a minor fraction of patients with ESCC.

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.

Odontogenic tumors: a retrospective study of four Brazilian diagnostic pathology centers.

OBJECTIVE: This article presents the results of a retrospective study of the frequency and classification of odontogenic tumors recorded at four centers of diagnostic pathology in Rio de Janeiro, Brazil. STUDY DESIGN: All medical records and microscopic slides of odontogenic tumor specimens for the years 1997 to 2007 were retrieved from the files of four services of diagnostic pathology in Rio de Janeiro City. Diagnoses were re-evaluated and the tumors classified according to the latest (2005) World Health Organization Classification of Tumors. RESULTS: A total of 201 odontogenic tumors were found among 15,758 oral biopsies (1.3%). The frequencies of these tumors at the four centers ranged from 0.5% at the National Cancer Institute to 3.3% in a private laboratory. Chi-square analysis revealed statistically significant differences (p<0.05) between the proportions of odontogenic tumors in the studied centers. Of these, 94.5% were benign and 5.5% were malignant. Keratocystic odontogenic tumor (32.3%) was the most frequent lesion, followed by ameloblastoma (29.8%) and odontoma (18.4%). CONCLUSIONS: Odontogenic tumors are uncommon in Brazil. Different pathology laboratories reported divergent frequencies of odontogenic tumors, which may reflect institutional specializations and the patient populations served.