Estudo genômico do tumor embrionário hepatoblastoma / Genomic study of the embryonal tumor hepatoblastoma

O hepatoblastoma, câncer de fígado mais comum na infância, é um tumor embrionário que se supõe surgir da interrupção da diferenciação hepática durante a embriogênese. O genoma deste tipo tumoral carrega poucas alterações somáticas, principalmente aneuploidias cromossômicas e mutações em CTNNB1. Essa relativa escassez de mutações somáticas representa um desafio à estratificação de risco dos pacientes e ao desenvolvimento de terapias direcionadas. Neste trabalho, investigamos por sequenciamento de exoma o espectro de mutações somáticas em um grupo de 10 hepatoblastomas, pareados com suas respectivas amostras germinativas, incluindo um caso de tumor congênito. Os dados genômicos revelaram que os hepatoblastomas tem número reduzido de mutações somáticas codificadoras não-sinônimas (média de ~6 variantes/tumor, com exclusão do caso congênito), totalizando 94 mutações (92 diferentes) nos 10 tumores, mapeadas em 87 genes. Apenas três genes apresentaram mutações detectadas em mais de uma amostra, CTNNB1, CX3CL1 e CEP164. As mutações foram validadas pelo sequenciamento de um painel composto pelos genes identificados no exoma, também utilizado para investigar estes genes em um grupo adicional de 12 tumores; apenas mutações em CTNNB1 foram detectadas neste grupo adicional. Mutações somáticas em CTNNB1 foram detectadas em ~54% do grupo estudado (22 hepatoblastomas): sete variantes patogênicas do tipo nucleotídeo único (SNV) ou indel foram identificadas em oito hepatoblastomas (~36%), uma delas nunca previamente descrita (A21_S33del); deleções intragênicas foram detectadas por sequenciamento Sanger em quatro outros tumores (~18%). A proteína ß-catenina foi avaliada por imunohistoquímica, apresentando translocação para o núcleo, o que indica ativação da via WNT; esse resultado também foi observado em tumores nos quais mutações em CTNNB1 não foram detectadas. O principal achado do estudo do exoma de hepatoblastomas foi a identificação de uma mutação somática recorrente no éxon 3 do gene CX3CL1 (A235G), observada em dois diferentes tumores. A análise de expressão gênica e proteica de CX3CL1 e de seu receptor CX3CR1 revelou aumento de expressão de CX3CL1 em hepatoblastomas; este resultado foi replicado em duas coortes independentes. O detalhamento da análise evidenciou um padrão bimodal: (a) linfócitos infiltrados em regiões tumorais de inflamação pós-quimioterapia eram negativos para essas proteínas, que deveriam estar expressas neste tipo celular em condições normais, enquanto as células tumorais as expressavam; (b) nas áreas de necrose tumoral pós-quimioterapia, houve detecção das proteínas CX3CL1/CX3CR1 nos linfócitos, mas não nas células tumorais. Em conjunto, estes resultados sugerem que a ativação da via CX3CL1/CX3CR1 ocorre em parte dos hepatoblastomas, independentemente da detecção de mutações, o que parece ser um achado relevante, potencialmente relacionado a inflamação e/ou resistência à quimioterapia. Adicionalmente, três assinaturas mutacionais foram detectadas nos hepatoblastomas, duas delas com predomínio das assinaturas do COSMIC, HB-S1 (COSMIC 1 e 6, presentes em todos os tipos de câncer) e HB-S2, com similaridades à assinatura COSMIC 29, relacionada apenas a carcinoma oral de células escamosas (gengivo-bucal) associado ao hábito de mascar tabaco; uma nova assinatura mutacional foi observada em um subconjunto de hepatoblastomas (HB-S3), com padrão inespecífico de pequeno aumento de mutações C>A. As assinaturas mutacionais já relatadas para câncer de fígado não foram evidentes nestes hepatoblastomas, sugerindo um processo mutacional diferente em sua origem. Por fim, análise de mutações germinativas no caso de hepatoblastoma congênito levou à identificação de variantes germinativas em genes de predisposição a câncer (BRCA1 e FAH), levantando a questão do papel da predisposição genética no desenvolvimento destes tumores embrionários (AU)

Hepatoblastoma, the most common liver cancer in infancy, is an embryonal tumor supposed to arise from differentiation impairment during embryogenesis. Hepatoblastomas genomes carry few somatic changes, mainly chromosomal aneuploidies and mutations in the CTNNB1 gene. This relative paucity of somatic mutations poses a challenge to risk stratification and development of targeted therapies. In this work, we investigated the burden of somatic mutations in a cohort of 10 hepatoblastomas paired with their respective germline samples, including a case of congenital tumor. Data revealed a low number of non-synonymous somatic coding mutations (mean of ~6 variants/tumor), totalizing 94 mutations in the 10 tumors, mapped in 87 genes; only three genes exhibited mutations detected in more than one sample, CTNNB1, CX3CL1 and CEP164. Target sequencing was used for validation and screening of the mutated genes in an additional group of 12 tumors; only CTNNB1 mutations were detected in this additional group. CTNNB1 mutations were detected in ~54% of the cohort (22 hepatoblastomas): seven single nucleotide variant or indel mutations were identified in eight hepatoblastomas (~36%), including the A21_S33del mutation, not previously reported; intragenic deletions were detected by Sanger sequencing in 4 tumors (~18%). The ß-catenin protein was evaluated by immunohistochemistry, presenting translocation to the nucleus, indicating activation of the WNT pathway; this result was also observed in tumors without CTNNB1 mutations. The main finding of the exome study was the identification of a recurrent somatic mutation in the exon 3 of the CX3CL1 gene (A235G) in two different hepatoblastomas. Gene expression and protein analysis of CX3CL1 and its receptor CX3CR1 revealed increased expression of CX3CL1 in hepatoblastomas, a result that was replicated in two independent cohorts. A bimodal pattern of expression was observed: (a) lymphocytes infiltrated in tumor regions of inflammation post-chemotherapy were negative for these proteins, which should be expressed in this cell type under normal conditions, while the tumor cells expressed them; (b) in areas of tumor necrosis after chemotherapy, CX3CL1/CX3CR1 proteins were detected in lymphocytes, but not in tumor cells. Taken together, these results suggest that activation of the CX3CL1/CX3CR1 pathway occurs in part of the hepatoblastomas, regardless of mutation detection, potentially related to inflammation and/or resistance to chemotherapy. Additionally, three mutational signatures were detected, two of them with a predominance of signatures of COSMIC, HB-S1 (COSMIC 1 and 6, present in all types of cancer) and HB-S2 (COSMIC 29 signature, related only to oral cell carcinoma gingival-buccal associated with the habit of chewing tobacco). A new mutational signature was observed in a subset of hepatoblastomas (HB-S3), with a non-specific pattern of small increase in C>A mutations. Mutational signatures already reported for liver cancer were not evident in these hepatoblastomas, suggesting a different mutational process. Finally, an exploration of germline mutations in the congenital hepatoblastoma led to the identification of variants in genes of cancer predisposition (BRCA1 and FAH), raising the question of the role of genetic predisposition in the development of these embryonal tumors (AU)

Carcinoma Embrionário de Mediastino Posterior com Metástase Pulmonar: Relato de Caso / Carcinoma Embrionario de Mediastino Posterior con Metástasis Pulmonar: Relato de Caso / Posterior Mediastinum Embryonal Carcinoma with Pulmonary Metastasis: Case Report

Introdução: Os carcinomas embrionários são os mais raros e, geralmente, se apresentam quando já estão associados com outros componentes de células germinativas. Possuem características clínicas e radiológicas similares aos tumores de saco vitelino. Relato do caso: Paciente G.A.S.L, sexo masculino, 30 anos, ex-tabagista e etilista. Iniciou com quadro de dores no hemitórax esquerdo em fevereiro de 2018 com extensão dos sintomas para a região da coluna torácica e lombar. Realizou tomografia de abdômen total e tórax, com resultado sugestivo de tumor de mediastino, nódulos pulmonares, material tecidual com densidade de partes moles no mediastino posterior e numerosas linfonodomegalias retroperitoneais, evoluiu com paraplegia de membros inferiores por compressão nervosa e com hipoestesia. Realizou biópsia de tumor de mediastino posterior à esquerda com resultado de neoplasia maligna epitelial e diagnóstico de carcinoma embrionário extragonodal pouco diferenciado. O paciente apresentou metástase pulmonar, confirmando que esses tumores frequentemente se infiltram nos órgãos adjacentes. A quimioterapia baseada em cisplatina é o tratamento padrão, levando à melhora da sobrevida em pacientes com esse tipo de tumor. Após a quimioterapia, houve diminuição do volume tumoral, porém, seguiu com a paraplegia de membros inferiores em razão da compressão nervosa. Conclusão: Este estudo relata o caso de um paciente jovem, com tumor raro de células germinativas e metástase pulmonar, que evoluiu clinicamente estável após tratamento específico com quimioterápicos. Por ainda haver uma escassa literatura acerca do tema, este estudo traz novas evidências e achados.

Introduction: Embryonal carcinomas are the rarest, and usually present when they are already associated with other components of germ cells. They have clinical and radiological features similar to yolk sac tumors. Case report: Patient G.A.S.L, male, 30 years old, former smoker and alcoholic. Initially, the patient reported pain in the left hemithorax in February 2018 with extension of the symptoms to the region of the thoracic and lumbar spine. A tomography of the total abdomen and chest was performed, with result suggestive of mediastinal tumor, pulmonary nodules, tissue material with soft tissue density in the posterior mediastinum and numerous retroperitoneal lymph node enlargement, which evolved with paraplegia of the lower limbs by nerve compression and hypoesthesia. It was conducted a biopsy of a mediastinal tumor posterior to the left with result of malignant epithelial neoplasia and diagnosis of extragonadal embryonic carcinoma very little differentiated. The patient presented pulmonary metastasis confirming that these tumors frequently infiltrate into the adjacent organs. Cisplatin-based chemotherapy is the standard treatment, leading to improved survival in patients with this type of tumor. After chemotherapy, the tumor volume decreased, but the patient continued with paraplegia of lower limbs due to nerve compression. Conclusion: This study reports the case of a young patient with a rare germ cell tumor and pulmonary metastasis who evolved clinically stable after specific chemotherapy treatment. Because there is still scarce literature on the subject, this study brings new evidences and findings.

Introducción: Los carcinomas embrionarios son los más raros y generalmente se presentan cuando ya están asociados con otros componentes de células germinativas. Se presentan características clínicas y radiológicas similares a los tumores de saco vitelino. Relato del caso: Paciente G.A.S.L, sexo masculino, 30 años, ex tabaquista y etilista. Se inició con cuadro de dolores en el hemitórax izquierdo en febrero de 2018 con extensión de los síntomas para la región de la columna torácica y lumbar. Se realizó una tomografía de abdomen total y tórax, con resultado sugestivo de tumor de mediastino, nódulos pulmonares, material tisular con densidad de partes blandas en el mediastino posterior y numerosas linfonodomegalias retroperitoneales, evolucionó con paraplejia de miembros inferiores por compresión nerviosa y con hipoestesia. Se realizó biopsia de tumor de mediastino posterior a la izquierda con resultado de neoplasia maligna epitelial y diagnóstico de carcinoma embrionario extra gonodal poco diferenciado. El paciente presentó metástasis pulmonar confirmando que estos tumores frecuentemente se infiltran en los órganos adyacentes. La quimioterapia basada en cisplatino es el tratamiento estándar, llevando a la mejora de la supervivencia en pacientes con este tipo de tumor. Después de la quimioterapia hubo la disminución del volumen tumoral, sin embargo, siguió con la paraplejia de miembros inferiores debido a la compresión nerviosa. Conclusión: Este estudio informa el caso de un paciente joven con un tumor raro de células germinales y metástasis pulmonar que evolucionó clínicamente estable después de un tratamiento de quimioterapia específico. Debido a que todavía hay poca literatura sobre el tema, este estudio aporta nuevas pruebas y hallazgos.

Embryonal Carcinoma of the Pineal Gland Developed in an Adolescent Boy with Klinefelter Syndrome / 임상소아혈액종양

Klinefelter syndrome (KS) is characterized by small testes, gynecomastia, tall stature, and hypergonadotropic hypogonadism. This condition is associated with extra X chromosomes. It is well known that these aneuploidies predispose individuals to the development of several cancers. Moreover, there are many case reports that show KS patients to have a higher relative risk for the development of malignancy. However, incracranial germ cell tumor (ICGCT) associated with KS is very uncommon. Herein, we report delayed diagnosis of KS in a 15-year-old boy with ICGCT, embryonal carcinoma of the pineal gland, after multimodality treatment in Korea.

Hematoma retroperitoneal como presentación de cáncer testicular / Retroperitoneal hematoma debut as testicular cancer

Resumen Introducción: El hematoma retroperitoneal (HR) es una enfermedad infrecuente con una elevada morbimortalidad, siendo complicado cuando se presenta con dolor y shock hipovolémico. Presentación del caso: Paciente del sexo masculino, de 20 años de edad, sin antecedentes mórbidos. Ingresa en Urgencias por dolor abdominal en el flanco izquierdo, irradiado a dorso y testículo ipsilateral, de 6 h de evolución, de inicio súbito e intensidad severa; el paciente está pálido, hemodinámicamente estable, sin signos de irritación peritoneal. Se solicita pielo-TC por sospecha de litiasis ureteral, que muestra un extenso HR, probable aneurisma aórtico roto. Una angio-TC informa HR adyacente y anterior a psoasilíaco izquierdo, de20 × 11 × 8,5 cm, volumen 972 cc, adenopatías retroperitoneales paraaórticas bilaterales sangrantes y múltiples nódulos pulmonares bilaterales indicativos de diseminación secundaria. Se constata testículo derecho duro, de tamaño normal, eco testicular con masa sólida quística, que indica de lesión orgánica. Discusión: Trauma y enfermedad tumoral son las principales causas de HR. El cáncer testicular suele presentarse en pacientes jóvenes, requiriendo una pronta derivación y estudio debido a su rápida progresión. En nuestro caso, el HR fue un hallazgo imagenológico, destacando que el sangrado de un conglomerado de adenopatías es anecdótico.

Abstract Introduction: Retroperitoneal hematoma (RH) is a rare disease with high morbidity, being complicated when presented with pain and hypovolemic shock. Case report: Male, 20 years old, no morbid history. Arrive to Emergency Service for abdominal pain in the left flank radiating to the back and ipsilateral testis, 6 h of evolution, sudden onset, high intensity; pacient pale, hemodynamically stable without signs of peritoneal irritation. Pielo-TC is requested on suspicion of ureteral stones showing extensive RH, likely ruptured aortic aneurysm. CT angiography reports RH and adjacent preceding left iliopsoas, 20 × 11 × 8.5 cm, volume 972 cc, retroperitoneal bleeding bilateral para-aortic lymphadenopathy and multiple bilateral pulmonary nodules suggestive of secondary spread. Hard right testicle with normal size, testicular ultrasound pointing solid cystic mass, suggestive of organic lesion. Discussion: Trauma and tumor pathology are the main causes of RH. Testicular cancer usually occurs in young patients, requiring early referal and study because of its rapid progression. In our case, the HR was an imaging finding, highlighting that the bleeding of a cluster of lymph nodes is anecdotal.

Pure Yolk sac presenting with inferior vena cava thrombus extending from bilateral external iliac veins to hepatic vein

ABSTRACT Introduction: Vena cava thrombus is an extremely rare complication of testicular tumors. We report on an unusual case of testicular tumor presenting with inferior vena cava thrombus extending from the left spermatic and bilateral external iliac veins to the hepatic vein. Case report: A-35-year old man presented with a 6-month history of left scrotal mass and a 1-day history of bilateral lower extremity edema. Computed tomography (CT) revealed the presence of thrombus extending from the left spermatic vein and bilateral external iliac veins to the hepatic vein, and multiple lymph node and lung metastases. 3 cycles of chemotherapy were given after the left high inguinal orchiectomy. Pathological examination demonstrated a pure yolk sac carcinoma with lymphovascular invasion and direct tumor extension into the left spermatic cord. CT and positron emission tompgraphy-CT obtained no findings of metastasis or recurrence at 3 months after the chemotherapy. Conclusion: We review this seldom case and discuss the literature with regard to its diagnosis and treatment.

Treatments of stage-Is testicular mixed germ cell tumors: A report of 3 cases / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate different treatment methods for stage-Is testicular mixed germ cell tumors (TMGCTs).</p><p><b>METHODS</b>We retrospectively analyzed the clinical data about 3'cases of stage-Is TMGCTs (aged 26-39 years) treated in the 175th Hospital of PLA, reviewed relevant literature, and explored the clinical characteristics of TMGCTs.</p><p><b>RESULTS</b>Of the 3 patients, 1 was treated by radical orchiectomy, 1 by radical orchiectomy + retroperitoneal lymph node dissection + BEP chemotherapy scheme, and the other by radical orchiectomy + radiotherapy. The pathological components of TMGCTs were immature teratoma, seminoma, spermatocytoma, chorioepithelioma, embryonal carcinoma, and yolk sac tumor. No recurrence or distant metastasis was found during the 24-month follow-up after surgery.</p><p><b>CONCLUSION</b>The diagnosis of TMGCTs primarily depends on physical examination, ultrasonography, MRI, and measurement of serum tumor markers, while its confirmation necessitates pathological examination, and its treatment is basically radical orchiectomy.</p>

Diagnostic value of immunohistochemistry and FISH for chromosome 12p in type Ⅱ testicular germ cell tumors / 中华男科学杂志

<p><b>Objective</b>To study the pathological morphology, immunohistochemical characteristics, and molecular changes of type Ⅱ testicular germ cell tumors (TGCT) and investigate the possible value of immunohistochemistry and fluorescence in situ hybridization (FISH) in the diagnosis of TGCT.</p><p><b>METHODS</b>We collected for this study 97 cases of TGCT, including 75 cases of seminoma, 17 cases of embryonal carcinoma, 11 cases of yolk sac tumor, 16 cases of mature teratoma, 3 cases of immature teratoma, and 1 case of epidermoid cyst, in which normal testicular tissue was found in 20 and non-TGCT in 6. We detected the expressions of different antibodies in various subtypes of TGCT by immunohistochemistry and determined the rate of chromosome 12p abnormality using FISH.</p><p><b>RESULTS</b>The immunophenotypes varied with different subtypes of TGCT. SALL4 and PLAP exhibited high sensitivity in all histological subtypes. CD117 and OCT4 showed strongly positive expressions in invasive seminoma and germ cell neoplasia in situ (GCNIS) but not in normal seminiferous tubules. GPC3 was significantly expressed in the yolk sac tumor, superior to GATA3 and AFP in both range and intensity. CKpan, OCT4, and CD30 were extensively expressed in embryonal carcinoma, while HCG expressed in choriocarcinoma. The positivity rate of isochromosome 12p and 12p amplification in TGCT was 96.7% (29/30).</p><p><b>CONCLUSIONS</b>The majority of TGCT can be diagnosed by histological observation, but immunohistochemical staining is crucial for more accurate subtypes and valuable for selection of individualized treatment options and evaluation of prognosis. Chromosome 12p abnormality is a specific molecular alteration in type Ⅱ TGCT, which is useful for ruling out other lesions.</p>

Testicular embryonal cell carcinoma presenting as haemoptysis and skin nodules.

Embryonal cell carcinoma affects young males in the prime of their life with majority of tumours already having metastasised at the time of diagnosis. Subcutaneous metastasis from embryonal carcinoma is rare and is associated with widespread disease and poor prognosis. We report a case of 22-year-old male who presented with haemoptysis and skin nodules. Fine needle aspiration cytology of skin nodules and the lung lesion led to the diognosis of testicular embryonal cell carcinoma.

Diagnostic value of MRI in testicular tumor / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the MRI manifestation of testicular tumor and the value of MRI in the diagnosis of the disease.</p><p><b>METHODS</b>We retrospectively analyzed 23 cases of pathologically confirmed testicular tumor, and observed the morphological characteristics, signals and surrounding conditions of the tumor using plain and enhanced MRI scanning.</p><p><b>RESULTS</b>Of the 23 cases, seminoma was identified in 7, mixed germinoma in 3, teratoma in 3, endodermal sinus tumor in 2, epidermoid in 1, Leydig cell tumor in 1, leucoma in 1, nonspecific inflammatory mass in 3, and tuberculosis in 2. MRI revealed the precise locations and specific characteristics of</p><p><b>CONCLUSION</b>Based on MRI findings and clinical manifestation, most testicular tumors can be diagnosed correctly.</p>

Antiproliferation and Redifferentiation in Thyroid Cancer Cell Lines by Polyphenol Phytochemicals

Thyroid carcinogenesis is accompanied by loss of thyroid-specific functions and refractory to radioiodine and thyroid stimulating hormone (TSH) suppression therapy. Redifferentiating agents have been shown to inhibit tumor growth and improve the response to conventional therapy. Polyphenol phytochemicals (PPs) in fruits and vegetables have been reported to inhibit cancer initiation, promotion, progression and induce redifferentiation in selected types. In this study we examined PPs induce redifferentiation in thyroid cancer cell lines. We investigated the effects of genistein, resveratrol, quercetin, kaempferol, and resorcinol on the F9 embryonal carcinoma cell differentiation model. The thyroid cancer cell lines, TPC-1, FTC-133, NPA, FRO, and ARO, displayed growth inhibition in response to genistein, resveratrol, quercetin. We further demonstrated that genistein decreased the dedifferention marker CD97 in NPA cells and resveratrol decreased CD97 in FTC-133, NPA, FRO cells and quercetin decreased CD97 in all cell lines. We observed increased expression of differentiation marker NIS in FTC-133 cells in response to genistein, and resveratrol but no change in NPA, FRO, ARO cells. Quercetin increased or induced NIS in FTC-133, NPA, FRO cells. These findings suggest that PPs may provide a useful therapeutic intervention in thyroid cancer redifferentiation therapy.

Letter to the Editor: Antiproliferation and Redifferentiation in Thyroid Cancer Cell Line by Polyphenol Phytochemicals

No abstract available.

Clinical features and microsurgical management of rare tumors in the sellar region / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the clinical characteristics and microsurgical managements of rare tumors in the sellar region.</p><p><b>METHODS</b>Six rare cases of tumors in the sellar region treated by microsurgery from Jan 2000 to Jan 2010 were reviewed retrospectively. Subsequent treatments were according to the status of preoperative alpha fetal protein (AFP) and human chorionic gonadotropin (HCG) measurement as well as confirmed by histopathological examination in all six patients.</p><p><b>RESULTS</b>Total resection of the tumor was achieved in 2 cases and subtotal resection in 4 cases. Postoperative histopathology confirmed that the lesions were tumors in 5 cases and fungal pseudotumor in 1 case. Moreover, variety of histological types were observed in the present series, including leiomyosarcoma, malignant yolk sac tumor, mixed germ cell tumor, embryonal carcinoma, pilocytic astrocytoma and fungal pseudotumor, respectively. The serum levels of AFP and HCG were elevated to some extent in the patients with malignant yolk sac tumor, mixed germ cell tumor or embryonal carcinoma. Follow-up was conducted in all patients for 1 month to 3 years. The patients with malignant yolk sac tumor and embryonal carcinoma as well as leiomyosarcoma died in 5, 6, 10 months after operation, respectively. Subarachnoid hemorrhage occurred in the case of fungal pseudotumor at 2 months after surgery. The other two patients were surviving well.</p><p><b>CONCLUSIONS</b>Rare non-germinomatous malignant germ cell tumors are predominantly susceptible to the sellar region. Furthermore, High misdiagnosis rate and poor prognosis are characteristic in the present study. Dynamic AFP and HCG detection may play an important role in the diagnosis of those non-germinomatous malignant germ cell tumors located in the sellar region. The importance of awareness of the presence of such rare lesions in the sellar region is emphasized.</p>

Intracardiac Metastasis of Testicular Embryonal CarcinomaThat Presented with a Right Ventricular Mass / 대한흉부외과학회지

Metastases to the heart are rarely diagnosed before the patient dies. A 26-year-old man was admitted with multiple metastasis of a testicular embryonal carcinoma and he was found to have intracardiac metastasis. Echocardiography showed that he had a mass rising from the interventricular septum and it was floating through the right ventricular outflow tract. The histology of the mass we removed from the right ventricle was consistent with testicular embryonal carcinoma. The patient made a smooth recovery after surgical intervention and chemotherapy. We believe this is the first reported case of testicular embryonal carcinoma that metastasized to the heart and that was successfully removed via surgery in Korea.

Enhanced Cardiomyogenic Differentiation of P19 Embryonal Carcinoma Stem Cells

BACKGROUND AND OBJECTIVES: We investigated the effects of different concentrations of serum, 5-azacytidine, and culture time on the cardiomyogenic differentiation of P19 embryonal carcinoma stem cells in the course of developing an efficient protocol for generating the cardiomyogenic lineage. MATERIALS AND METHODS: P19 cells were plated at a density of 1x10(6) cells on 10-cm bacterial dishes for 96 hours in the presence of 1% dimethyl sulfoxide to form embryoid bodies. The embryoid bodies were cultured in medium with 2% or 10% fetal bovine serum for an additional 10 or 15 consecutive days in the presence of 0, 1, or 3 microM 5-azacytidine. RESULTS: Quantitative real-time polymerase chain reaction (PCR) analysis showed that the messenger ribonucleic acid (mRNA) expression of cardiac muscle-specific genes, such as GATA4, alpha-actin, alpha-myosin heavy chain, and cardiac troponin T, were significantly higher in the 15-day culture groups than in the 10-day culture groups. Furthermore, the cardiac muscle-specific genes were expressed more in the high-serum groups compared to the low-serum groups regardless of the culture time. Cardiomyogenic differentiation of the P19 cells was most effective in 1 microM 5-azacytidine regardless of the serum concentrations. In addition, the stimulation effects of 5-azacytidine on cardiomyogenic differentiation were more significant under low-serum culture conditions compared to high-serum culture conditions. Cardiomyogenic differentiation of P19 cells was further confirmed by immunostaining with cardiac muscle-specific antibodies. CONCLUSION:Taken together, these results demonstrated that cardiomyogenic differentiation of P19 cells was enhanced by a combination of different experimental factors.

Identification of human testicular embryonal carcinoma proteins by two-dimensional gel electrophoresis and mass spectrometry / 南方医科大学学报

<p><b>OBJECTIVE</b>To separate and identify human testicular embryonal carcinoma proteomics using two-dimensional electrophoresis (2-DE) and mass spectrometry.</p><p><b>METHODS</b>Immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis was used to separate the total proteins of the samples. After silver staining, PDQuest 7.30 image analysis software was applied to analyze the 2-DE images. Three of the proteins highly expressed in human testicular embryonal carcinoma were identified by matrix-assisted laser adsorption/ionization-time of flight-tandem mass spectrometry (MALDI-TOF-MS/MS).</p><p><b>RESULTS</b>2-DE effectively screened the differentially expressed proteins in the carcinoma tissues. Three proteins highly expressed in the carcinoma were successfully identified.</p><p><b>CONCLUSION</b>The proteins of human testicular embryonal carcinoma can be effectively separated and analyzed using 2-DE and mass spectrometry. Proteomic analysis offers a new means for further study of this carcinoma.</p>

Tumor de células germinales / Germ cell tumour / Tumor de células germinativas

Los tumores de células germinales son neoplasias que se derivan de las células germinales primordiales. Histológicamente se clasifican como tumores seminomatosos o no seminomatosos, dividiéndose estos últimos, según la diferenciación celular, en carcinoma embrionario, coriocarcinomas, tumor del saco vitelino y teratoma maduro e inmaduro. Se trata de los carcinomas sólidos más frecuentes en varones entre 20 y 35 años y corresponden al 90% de los tumores testiculares, cursando en testículo como una masa de consistencia dura, usualmente asintomática y no dolorosa. Su origen embrionario, así como la migración desde el intestino primordial hasta los túbulos seminíferos, explican la aparición de tumores de células germinales mediastinales y retroperitoneales. En la presente revisión se hace la descripción de las características embriológicas, patológicas y clínicas de la enfermedad, así como su estadificación y tratamiento...

Germ Cell Tumours are malignancies derived from primordial germ cells. They are classified as seminomatous and non seminomatous, the latter divided based on the cellular differentiation as Embrionary germ cell, Choriocarcinomas, Yolk salk tumors and mature and inmature teratoma. They are the most frequent tumors in men between the ages of 20 and 35 and correspond to 90% of testicular tumors found as non-symptomatic hard painless testicular masses. Their embryologic origin as well as their migration from the primordial gut toward the seminiferous tubules explains the emergence of GCT in the mediastinum and retroperitoneum. In this review there is a description about embryologic, clinical and pathologic characteristics of the disease, as well as staging and treatment...

Os tumores de células germinais são neoplasias derivadas das células germinais primordiais. Classificam-se como tumores seminomatosos ou não seminomatosos, dividindo-se estes últimos, segundo a diferenciação celular, em carcinoma embrionário, coriocarcinomas, tumor do saco vitelino e teratoma maduro e imaturo. Trata-se dos carcinomas sólidos mais freqüentes em varões entre 20 e 35 anos e correspondem ao 90% dos tumores testiculares, cursando em testículo como uma massa de consistência dura, usualmente assintomática e não dolorosa. Sua origem embriológica, bem como a migração desde o intestino primordial até os túbulos seminíferos, explica a aparição de tumores de células germinais mediastinais e retroperitonais. Na presente revisão se faz a descrição das características embriológicas, patológicas e clínicas da doença, bem como sua estadifição e tratamento...

Outcome of patients with stage II and III nonseminomatous germ cell tumors: results of a single center.

BACKGROUND: The prognostic factors in nonseminomatous germ cell tumors have been mainly derived from the analysis of stage I tumors. AIMS: The aim of this study was to evaluate some prognostic factors and the outcome of patients with stage II and III nonseminomatous germ cell tumors according to risk groups treated between 1993 and 2002. SETTINGS AND DESIGN: Patients were retrospectively classified as good, intermediate and poor risk groups according to International Germ Cell Cancer Consensus Group. MATERIALS AND METHODS: Biopsy specimens of 58 patients with stage II and III nonseminomatous germ cell tumors were analyzed by means of tumor histopathology, primary localization site of the tumor, relapse sites, initial serum tumor marker levels, the presence of persistent serum tumor marker elevation and the patients' outcome. STATISTICAL ANALYSIS: Kruskall Wallis test and Mann-Whitney U test were used to determine the differences between the groups. Kaplan-Meier method was used for survival analysis and log rank test was used to compare the survival probabilities of groups. Cox proportional hazard analysis was used to determine the prognostic factors in univariate and multivariate analysis. RESULTS: Five-year overall and disease-free survival rates were calculated as 85% and 75% in stage II; 44% and 29% in stage III cases, respectively. Fifty-seven percent of patients were classified in good risk, 9% in intermediate risk and 27% in poor risk groups. Five-year overall survival rates were 97%, 75% and 7% (P<0.001) and disease-free survival rates were 83%, 34% and 7% (P<0.001) in good, intermediate and poor risk groups, respectively. Analysis of the prognostic factors revealed that the localization site of the primary tumor (P<0.001), the initial beta-HCG level (p:0.0048), the presence of yolk sac and choriocarcinoma components in tumor (p:0.003 and p:0.004), relapse sites of tumor (lung versus other than lung) (p:0.003), persistent elevation of serum tumor markers (P<0.001) were significant prognostic factors in univariate analysis. However, in multivariate analysis, only the localization site of tumor (p:0.049) and the relapse site (p:0.003) were found statistically significant. CONCLUSIONS: This retrospective study revealed that in advanced stage of nonseminomatous germ cell tumors, the outcome is essentially related with the localization site of the tumor and the relapse site.

Pure Immature Teratoma with Increase of Serum alpha-fetoprotein: A Case Report

In the immature teratoma with increased serum alpha-fetoprotein (AFP), a complete pathologic examination is especially required because coexistence with other germ cell tumors is associated with a poor prognosis. If the case is proved to be a pure immature teratoma in spite of a thorough examination, the source of AFP should be found as an AFP-producing pure immature teratoma is not associated with a poor prognosis. In this case of a grade III-immature teratoma in an ovary of a 12-year old girl, serum AFP was increased. On pathologic examination, there was no evidence of a yolk sac tumor or embryonal carcinoma. On an AFP immunohistochemical stain, immature liver tissue, digestive and respiratory epitheliums were positive.