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1.
Int J Mol Sci ; 22(5)2021 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-33800799

RESUMO

Liquid biopsies constitute a minimally invasive means of managing cancer patients, entailing early diagnosis, follow-up and prediction of response to therapy. Their use in the germ cell tumor field is invaluable since diagnostic tissue biopsies (which are invasive) are often not performed, and therefore only a presumptive diagnosis can be made, confirmed upon examination of the surgical specimen. Herein, we provide an overall review of the current liquid biopsy-based biomarkers of this disease, including the classical, routinely used serum tumor markers-the promising microRNAs rapidly approaching the introduction into clinical practice-but also cell-free DNA markers (including DNA methylation) and circulating tumor cells. Finally, and importantly, we also explore novel strategies and challenges for liquid biopsy markers and methodologies, providing a critical view of the future directions for liquid biopsy tests in this field, highlighting gaps and unanswered questions.


Assuntos
Biópsia Líquida , Neoplasias Embrionárias de Células Germinativas/diagnóstico , Biomarcadores Tumorais/análise , Biomarcadores Tumorais/sangue , Procedimentos Clínicos , DNA de Neoplasias/química , Gerenciamento Clínico , Feminino , Humanos , Masculino , MicroRNAs/análise , Proteínas de Neoplasias/análise , Neoplasias Embrionárias de Células Germinativas/sangue , Neoplasias Embrionárias de Células Germinativas/química , Neoplasias Embrionárias de Células Germinativas/patologia , Células Neoplásicas Circulantes , Neoplasias Ovarianas/sangue , Neoplasias Ovarianas/química , Neoplasias Ovarianas/diagnóstico , Neoplasias Ovarianas/patologia , RNA Neoplásico/análise , Neoplasias Testiculares/sangue , Neoplasias Testiculares/química , Neoplasias Testiculares/diagnóstico , Neoplasias Testiculares/patologia
2.
Int J Mol Sci ; 22(4)2021 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-33671303

RESUMO

In the present era, infertility is one of the major issues which restricts many couples to have their own children. Infertility is the inability to achieve a clinical pregnancy after regular unprotected sexual intercourse for the period of one year or more. Various factors including defective male or female germ cell development, unhealthy and improper lifestyles, diseases like cancer and associated chemo-or-radiation therapies, congenital disorders, etc., may be responsible for infertility. Therefore, it is highly important to understand the basic concepts of germ cell development including primordial germ cell (PGC) formation, specification, migration, entry to genital ridges and their molecular mechanisms, activated pathways, paracrine and autocrine signaling, along with possible alteration which can hamper germ cell development and can cause adversities like cancer progression and infertility. Knowing all these aspects in a proper way can be very much helpful in improving our understanding about gametogenesis and finding possible ways to cure related disorders. Here in this review, various aspects of gametogenesis especially female gametes and relevant factors causing functional impairment have been thoroughly discussed.


Assuntos
Células Germinativas/patologia , Animais , Carcinogênese/patologia , Epigênese Genética , Feminino , Humanos , Neoplasias Embrionárias de Células Germinativas/complicações , Neoplasias Embrionárias de Células Germinativas/diagnóstico , Neoplasias Embrionárias de Células Germinativas/genética , Neoplasias Embrionárias de Células Germinativas/patologia , Oócitos/citologia
3.
Biomed Environ Sci ; 34(2): 152-162, 2021 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-33685574

RESUMO

Objective: Testicular germ cell tumors (TGCT) are the most common cancer among men aged 15 to 39 years. Previous studies have considered factors related to TGCT survival rate and race/ethnicity, but histological type of the diagnosed cancer has not yet been thoroughly assessed. Methods: The data came from 42,854 eligible patients from 1992 to 2015 in the Surveillance Epidemiology and End Results 18. Frequencies and column percent by seminoma and nonseminoma subtypes were determined for each covariates. We used Cox proportional hazard regression to assess the impact of multiple factors on post-diagnostic mortality of TGCT. Results: Black males were diagnosed at a later stage, more commonly with local or distant metastases. The incidence of TGCT in black non-seminoma tumors increased most significantly. The difference in survival rates between different ethnic and histological subtypes, overall survival (OS) in patients with non-seminoma was significantly worse than in patients with seminoma. The most important quantitative predictor of death was the stage at the time of diagnosis, and older diagnostic age is also important factor affecting mortality. Conclusion: Histological type of testicular germ cell tumor is an important factor in determining the prognosis of testicular cancer in males of different ethnic groups.


Assuntos
Disparidades nos Níveis de Saúde , Neoplasias Embrionárias de Células Germinativas/etnologia , Neoplasias Embrionárias de Células Germinativas/mortalidade , Neoplasias Testiculares/etnologia , Neoplasias Testiculares/mortalidade , Adulto , Humanos , Masculino , Neoplasias Embrionárias de Células Germinativas/diagnóstico , Neoplasias Embrionárias de Células Germinativas/patologia , Prognóstico , Fatores de Risco , Programa de SEER/estatística & dados numéricos , Seminoma/diagnóstico , Seminoma/etnologia , Seminoma/mortalidade , Seminoma/patologia , Taxa de Sobrevida/tendências , Neoplasias Testiculares/diagnóstico , Neoplasias Testiculares/patologia , Estados Unidos/epidemiologia , Estados Unidos/etnologia
4.
Eur J Endocrinol ; 184(4): 617-625, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33524006

RESUMO

Objective: To investigate the incidence of hypothalamus-pituitary-gonadal (HPG) axis initiation/recovery after treatment and to identify predictive risk factors for noninitiation/recovery. Methods: A total of 127 consecutive suprasellar germ cell tumor (GCT) patients managed at Peking Union Medical College Hospital (2006-2019) were retrospectively analyzed. Prepubertal patients (followed up until 13 years of age for girls and 14 years of age for boys) and patients with HPG dysfunction (followed up for 2 years) were divided into the initiation/recovery and noninitiation/recovery groups. Results: Of the 127 suprasellar GCT patients, 75 met the follow-up criteria, 28 (37.3%) of whom experienced HPG axis initiation/recovery. Compared to the noninitiation/recovery group, the initiation/recovery group included more males and had shorter delayed diagnosis times, smaller tumor sizes, lower panhypopituitarism rates, thinner pituitary stalk widths, lower visual deficit rates, and higher serum testosterone and estradiol levels. The cutoff values of pituitary stalk width, tumor size, and delayed diagnosis time used to predict noninitiation/recovery were 6.9 mm, 6.9 mm and 1.7 years, respectively. Tumor size ≥6.9 mm (odds ratio (OR) = 7.5, 95% CI: 2.2-25.8, P = 0.001), panhypopituitarism (OR = 5.0, 95% CI: 1.4-17.6, P = 0.013), and delayed diagnosis time ≥1.7 years (OR = 5.7, 95% CI: 1.5-20.7, P = 0.009) were risk factors for noninitiation/recovery. Conclusions: Among suprasellar GCT patients, nearly one-third of prepubertal patients and patients with HPG dysfunction experience HPG axis initiation/recovery after treatment. Tumor size ≥6.9 mm, panhypopituitarism, and delayed diagnosis time ≥1.7 years were identified as predictive risk factors for noninitiation/recovery.


Assuntos
Gônadas/fisiologia , Sistema Hipotálamo-Hipofisário/fisiologia , Neoplasias Embrionárias de Células Germinativas/terapia , Neoplasias Hipofisárias/terapia , Recuperação de Função Fisiológica/fisiologia , Adolescente , Idade de Início , Estudos de Casos e Controles , Criança , China/epidemiologia , Feminino , Hormônio Foliculoestimulante/sangue , Seguimentos , Humanos , Hipotálamo/fisiologia , Hormônio Luteinizante/sangue , Masculino , Neoplasias Embrionárias de Células Germinativas/diagnóstico , Neoplasias Embrionárias de Células Germinativas/epidemiologia , Neoplasias Embrionárias de Células Germinativas/reabilitação , Neoplasias Hipofisárias/diagnóstico , Neoplasias Hipofisárias/epidemiologia , Neoplasias Hipofisárias/reabilitação , Prognóstico , Puberdade/sangue , Puberdade/fisiologia , Estudos Retrospectivos , Testosterona/sangue
5.
Crit Rev Oncol Hematol ; 159: 103224, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33493632

RESUMO

Germ cell tumors (GCTs) represent the best and the only example of solid tumors curable in the large majority of patients. GCTs are one of the few malignancies for which specific biochemical markers have been identified: human chorionic gonadotropin (HCG) and alfa-fetoprotein (AFP). Due to their specificity and sensitivity they constitute formidable tools in the diagnosis and monitoring of treatment for GCTs. As a tumor mass marker, lactate dehydrogenase (LDH) is also considered. Tumor markers are expressed in 15-20% of seminoma and 60-80% of non-seminoma. With the aim to increase sensitivity and specificity, recent studies have proposed miRNAs as serum biomarkers. This review will focus on role of serum tumor markers in diagnosis, staging, prognosis, monitoring of response, and finally follow-up of GCTs.


Assuntos
Neoplasias Embrionárias de Células Germinativas , Seminoma , Neoplasias Testiculares , Biomarcadores Tumorais , Humanos , Masculino , Neoplasias Embrionárias de Células Germinativas/diagnóstico , Neoplasias Embrionárias de Células Germinativas/terapia , Prognóstico , Neoplasias Testiculares/diagnóstico , Neoplasias Testiculares/terapia
6.
Am J Surg Pathol ; 45(1): 127-136, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-32991342

RESUMO

Rare hepatoid teratomas (HTs) in testicular germ cell tumor patients mimic hepatoid yolk sac tumor (HYST) and hepatocellular carcinoma (HCC). We compared the features of 2 metastatic HTs, 12 HYSTs, and 16 HCCs. The mean ages were 36, 40, and 62.5 years, respectively. The HTs formed sheets of hepatocyte-like cells with macrovesicular fat arranged in vague lobules with intervening fibrous bands containing biliary ductule-like structures and abortive portal triads. HTs lacked basement membrane deposits, with hepatoid cells staining for glypican-3, arginase, and HepPar-1 (2/2), whereas stains for CK19 (2/2) and CK7 (1/2) highlighted ductules and for villin hepatoid cells and ductules (1/2). SALL4 and CDX2 stains were negative (0/2). HYSTs formed nests, trabeculae, cords, and occasional gland-like structures, and most (10/12; 83%) produced intercellular basement membrane. No Mallory-Denk bodies were seen. Stains for SALL4 (100%), glypican-3 (100%), CK19 (88%), CDX2 (88%), and villin (75%) were positive, whereas those for HepPar-1 highlighted rare tumor cells (70%) and for arginase were mostly negative (26%). All HCCs lacked basement membrane deposits, with Mallory-Denk bodies occurring in 50%. Stains for HepPar-1 (100%) and arginase (94%) were positive, glypican-3 infrequent (19%), and SALL4, CK19, villin, and CDX2 negative. In summary, HTs are distinguished from HYST by the formation of ductules and abortive portal tracts, lack of basement membrane deposits, more consistent staining for arginase and HepPar-1, and negativity for SALL4 and CDX2. Contrasting features of HCCs with HYSTs include negativity for SALL4, CK19, and CDX2, frequent Mallory-Denk bodies, and absence of basement membrane deposits.


Assuntos
Carcinoma Hepatocelular/patologia , Neoplasias Hepáticas/patologia , Neoplasias Embrionárias de Células Germinativas/patologia , Teratoma/patologia , Neoplasias Testiculares/patologia , Adolescente , Adulto , Carcinoma Hepatocelular/diagnóstico , Diagnóstico Diferencial , Tumor do Seio Endodérmico , Humanos , Imuno-Histoquímica , Neoplasias Hepáticas/diagnóstico , Masculino , Pessoa de Meia-Idade , Neoplasias Embrionárias de Células Germinativas/diagnóstico , Teratoma/diagnóstico , Neoplasias Testiculares/diagnóstico , Adulto Jovem
7.
J Urol ; 205(1): 137-144, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32856980

RESUMO

PURPOSE: Current serum tumor markers for testicular germ cell tumor are limited by low sensitivity. Growing evidence supports the use of circulating miR-371a-3p as a superior marker for malignant (viable) germ cell tumor management. We evaluated the real-world application of serum miR-371a-3p levels in detecting viable germ cell tumor among patients undergoing partial or radical orchiectomy. MATERIALS AND METHODS: Serum samples were collected from 69 consecutive patients before orchiectomy. Performance characteristics of serum miR-371a-3p were compared with conventional serum tumor markers (⍺-fetoprotein/ß-human chorionic gonadotropin/lactate dehydrogenase) between patients with viable germ cell tumor and those without viable germ cell tumor on orchiectomy pathology. Relative miR-371a-3p levels were correlated with clinical course. The Kruskal-Wallis test and linear and ordinal regression models were used for analysis. RESULTS: For detecting viable germ cell tumor, combined conventional serum tumor markers had a specificity of 100%, sensitivity of 58% and AUC of 0.79. The miR-371a-3p test showed a specificity of 100%, sensitivity of 93% and AUC of 0.978. Median relative expression of miR-371a-3p in viable germ cell tumor cases was more than 6,800-fold higher than in those lacking viable germ cell tumor. miR-371a-3p levels correlated with composite stage (p=0.006) and, among composite stage I cases, independently associated with embryonal carcinoma percentage (p=0.0012) and tumor diameter (p <0.0001). Six patients underwent orchiectomy after chemotherapy and were correctly predicted to have presence or absence of viable germ cell tumor by the miR-371a-3p test. CONCLUSIONS: If validated, the miR-371a-3p test can be used in conjunction with conventional serum tumor markers to aid clinical decision making. A positive miR-371a-3p test in patients after preoperative chemotherapy or with solitary testes could potentially guide subsequent orchiectomy or observation.


Assuntos
Biomarcadores Tumorais/sangue , MicroRNA Circulante/sangue , MicroRNAs/sangue , Neoplasias Embrionárias de Células Germinativas/diagnóstico , Orquiectomia , Neoplasias Testiculares/diagnóstico , Adulto , Estudos de Casos e Controles , Quimioterapia Adjuvante , Tomada de Decisão Clínica/métodos , Estudos de Viabilidade , Humanos , Masculino , Pessoa de Meia-Idade , Terapia Neoadjuvante/métodos , Estadiamento de Neoplasias , Neoplasias Embrionárias de Células Germinativas/sangue , Neoplasias Embrionárias de Células Germinativas/patologia , Neoplasias Embrionárias de Células Germinativas/terapia , Período Pré-Operatório , Neoplasias Testiculares/sangue , Neoplasias Testiculares/patologia , Neoplasias Testiculares/terapia , Testículo/patologia , Testículo/cirurgia , Conduta Expectante
8.
Methods Mol Biol ; 2195: 1-11, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32852753

RESUMO

This chapter introduces the macroscopic and light microscopic features of testicular germ cell tumors (GCT) commonly encountered in clinical practice. Accurate diagnosis of these histologically diverse neoplasms is essential not only for clinical management but also for serving as the basis for interpretation of research findings. We will focus on general histopathologic concepts and discuss the use of immunohistochemistry (IHC) as an aid to the diagnosis.


Assuntos
Gradação de Tumores , Estadiamento de Neoplasias , Neoplasias Embrionárias de Células Germinativas/diagnóstico , Biomarcadores Tumorais , Biópsia , Carcinoma in Situ , Tomada de Decisão Clínica , Diagnóstico Diferencial , Gerenciamento Clínico , Feminino , Humanos , Imuno-Histoquímica/métodos , Masculino , Microscopia/métodos , Gradação de Tumores/métodos , Estadiamento de Neoplasias/métodos , Pesquisa , Neoplasias Testiculares/diagnóstico
9.
Methods Mol Biol ; 2195: 13-29, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32852754

RESUMO

Germ cell tumors (GCT) in men comprise of tumor subtypes with distinct histomorphologies, genetic and genomic alterations, and clinical behavior. Immunohistochemical (IHC) markers, including many newly described nuclear transcription factors, are often applied in challenging cases to arrive at a correct diagnosis and classification, and to establish germ cell origin for metastatic tumors. However, there is no established role for IHC markers in prognosis and therapy response prediction in GCTs. This chapter briefly reviews the clinical utility of IHC in diagnosis and classification of GCTs, including technical aspects of performing IHC and clinical applications of commonly used IHC markers in the workup of common and clinically relevant diagnostic scenarios.


Assuntos
Imuno-Histoquímica , Neoplasias Embrionárias de Células Germinativas/diagnóstico , Biomarcadores Tumorais , Tomada de Decisão Clínica , Diagnóstico Diferencial , Gerenciamento Clínico , Humanos , Imuno-Histoquímica/métodos , Masculino , Neoplasias Embrionárias de Células Germinativas/etiologia , Prognóstico , Fatores Sexuais
10.
Methods Mol Biol ; 2195: 31-47, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32852755

RESUMO

Testicular germ cell tumors are among the most common malignancies seen in children and young adults. Genomic studies have identified characteristic molecular profiles in testicular cancer, which are associated with histologic subtypes and may predict clinical behavior including treatment responses. Emerging molecular technologies analyzing tumor genomics, transcriptomics, and proteomics may now guide precision management of testicular tumors. Laser-assisted microdissection methods such as laser capture microdissection efficiently isolate selected tumor cells from routine pathology specimens, avoiding contamination from nontarget cell populations. Laser capture microdissection in combination with next generation sequencing makes precise high throughput genetic evaluation effective and efficient. The use of laser capture microdissection (LCM) for molecular testing may translate into great benefits for the clinical management of patients with testicular cancers. This review discusses application protocols for laser-assisted microdissection to investigate testicular germ cell tumors.


Assuntos
Biomarcadores Tumorais , Microdissecção , Técnicas de Diagnóstico Molecular , Neoplasias Embrionárias de Células Germinativas/diagnóstico , Neoplasias Embrionárias de Células Germinativas/etiologia , Neoplasias Testiculares/diagnóstico , Neoplasias Testiculares/etiologia , Tomada de Decisão Clínica , Diagnóstico Diferencial , Gerenciamento Clínico , Suscetibilidade a Doenças , Humanos , Imuno-Histoquímica/instrumentação , Imuno-Histoquímica/métodos , Masculino , Microdissecção/instrumentação , Microdissecção/métodos , Técnicas de Diagnóstico Molecular/métodos
11.
Methods Mol Biol ; 2195: 49-63, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32852756

RESUMO

Gains of genetic material or internal rearrangements of chromosome 12p, including 12p overrepresentation or isochromosome 12p [i(12p)], are observed in virtually all germ cell tumors (GCT), in all histologic subtypes, and from various body locations. The chromosomal region involved in these alterations contains the growth and survival promoting oncogene KRAS (12p12.1). Gains or rearrangements of 12p characterize GCT from in situ to chemoresistant stages. Fluorescence in situ hybridization (FISH) detection of chromosome 12p anomalies is a sensitive and specific test for the diagnosis of germ cell tumors. Here we provide a detailed protocol for FISH detection of isochromosome 12p and chromosome 12p overrepresentation. The method is helpful for diagnosis of germ cell origin, and for selection of patients who may benefit from cisplatin-based chemotherapy.


Assuntos
Aberrações Cromossômicas , Cromossomos Humanos Par 12 , Hibridização in Situ Fluorescente , Neoplasias Embrionárias de Células Germinativas/diagnóstico , Neoplasias Embrionárias de Células Germinativas/genética , Neoplasias Testiculares/diagnóstico , Neoplasias Testiculares/genética , Biomarcadores Tumorais , Diagnóstico Diferencial , Estudos de Associação Genética , Predisposição Genética para Doença , Humanos , Imuno-Histoquímica , Hibridização in Situ Fluorescente/métodos , Hibridização in Situ Fluorescente/normas , Masculino , Controle de Qualidade
12.
Methods Mol Biol ; 2195: 65-76, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32852757

RESUMO

Optimization of cell culture protocol for a given cell line is critical for the proper conduct of in vitro experiments. Because germ cell tumors can be so heterogeneous, optimal culture conditions can vary widely between cell lines. Here, we describe our experience in routine culture and cryopreservation of germ cell tumor cell culture. Additionally, methods for measuring cell viability and proliferation validated on these lines are provided.


Assuntos
Técnicas de Cultura de Células , Neoplasias Embrionárias de Células Germinativas/patologia , Cultura Primária de Células , Linhagem Celular Tumoral , Sobrevivência Celular , Criopreservação/métodos , Humanos , Neoplasias Embrionárias de Células Germinativas/diagnóstico
13.
Methods Mol Biol ; 2195: 113-123, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32852761

RESUMO

Testicular germ cell tumors (TGCTs) are typically exquisitely sensitive to DNA interstrand cross-link (ICLs) agents. ICLs covalently link both strands of the DNA duplex, impeding fundamental cellular processes like DNA replication to cause cell death. A leading drug used for the treatment of TGCTs is cisplatin, which introduces ICLs and leads to formation of double strand breaks (DSBs), a DNA lesion that can be repaired in the S/G2 phases of the cell cycle by homologous recombination (HR, also termed homology-direct repair). Although most TGCTs respond to cisplatin-induced ICLs, a fraction is resistant to treatment. One proposed mechanism of TGCT resistance to cisplatin is an enhanced ability to repair DSBs by HR. Other than HR, repair of the ICL-lesions requires additional DNA repair mechanisms, whose action might also be implemented in therapy-resistant cells. This chapter describes GFP assays to measure (a) HR proficiency following formation of a DSB by the endonuclease I-SceI, and (b) HR repair induced by site-specific ICL formation involving psoralen. These experimental approaches can be used to determine the proficiency of TGCT cell lines in DSB repair by HR in comparison to HR repair of ICLs, providing tools to better characterize their recombination profile. Protocols of these assays have been adapted for use in Embryonal Carcinoma (EC) TGCT cell lines. Assays only require transient introduction of plasmids within cells, affording the advantage of testing multiple cell lines in a relatively short time.


Assuntos
Testes Genéticos , Recombinação Homóloga , Neoplasias Embrionárias de Células Germinativas/genética , Reparo de DNA por Recombinação , Neoplasias Testiculares/genética , Técnicas de Cultura de Células , Linhagem Celular Tumoral , Dano ao DNA , Replicação do DNA , Ficusina , Expressão Gênica , Genes Reporter , Testes Genéticos/métodos , Humanos , Masculino , Neoplasias Embrionárias de Células Germinativas/diagnóstico , Neoplasias Testiculares/diagnóstico
14.
Methods Mol Biol ; 2195: 189-223, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32852766

RESUMO

Genomewide association studies (GWAS) have been widely used in recent years to identify common variants that are associated with multiple types of cancer, including testicular germ cell tumors. These studies require no a priori hypotheses and have advantages, including the ability to highlight new pathways relevant to the biology of common diseases. GWAS require collection of germline DNA from individuals with and without the disease of interest. Following DNA extraction and quantification, a variety of array based platforms are available to evaluate common and moderately rare germline variation throughout the genome in an agnostic fashion. Here, we describe DNA extraction methods from samples typically used in the evaluation of germline genetic variation (blood and saliva). We also describe assays used to assess DNA quality and quantity. Finally, we include methods describing array based genotyping using the Illumina platform and validation of relevant variants using the iPLEX Agena Multiplexed Genotyping (formerly Sequenom).


Assuntos
Predisposição Genética para Doença , Estudo de Associação Genômica Ampla , Neoplasias Embrionárias de Células Germinativas/genética , Neoplasias Testiculares/genética , Alelos , Biomarcadores Tumorais , Estudo de Associação Genômica Ampla/métodos , Genótipo , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Biópsia Líquida/métodos , Masculino , Neoplasias Embrionárias de Células Germinativas/diagnóstico , Neoplasias Embrionárias de Células Germinativas/epidemiologia , Saliva/metabolismo , Neoplasias Testiculares/diagnóstico , Neoplasias Testiculares/epidemiologia
15.
Methods Mol Biol ; 2195: 225-243, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32852767

RESUMO

Reverse transcription (RT) based quantitative PCR (qPCR) for quantifying microRNAs (miRNAs) in in the circulation presents specific challenges. Here, we describe an optimized research protocol to assess serum sample quality and quantify levels of a panel of four test miRNAs (miR-371a-3p, miR-372-3p, miR-373-3p, and miR-367-3p) that enables highly sensitive and specific malignant germ cell tumor (GCT) diagnosis and monitoring. This protocol utilizes a multiplex RT step using Taqman miRNA stem-loop primers. A multiplexed preamplification stage is then employed to increase the sensitivity of the final quantification step, which is performed using standard singleplex Taqman qPCR methodology.


Assuntos
Biomarcadores Tumorais , MicroRNA Circulante , Neoplasias Embrionárias de Células Germinativas/diagnóstico , Neoplasias Embrionárias de Células Germinativas/genética , Interpretação Estatística de Dados , Humanos , Reação em Cadeia da Polimerase Multiplex , Neoplasias Embrionárias de Células Germinativas/sangue , Técnicas de Amplificação de Ácido Nucleico , Prognóstico , Controle de Qualidade , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reação em Cadeia da Polimerase em Tempo Real/normas , Fluxo de Trabalho
16.
Methods Mol Biol ; 2195: 245-261, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32852768

RESUMO

While the majority of patients with advanced testicular germ cell tumors (GCT) achieve complete responses after chemotherapy and if indicated after postchemotherapy resection of residual lesions, about 20% of patients have incomplete responses or show relapses. Moreover, toxicity of chemotherapy is high, and severe adverse chronic effects have been described. Therefore, there is an urgent need for biomarkers that could help to improve tumor staging, and support decision-making, ideally including monitoring of therapy response and prediction of relapse. Besides the well-established serum markers lactate dehydrogenase, α-fetoprotein, and ß-subunit of human chorionic gonadotropin, during recent years new noninvasive liquid biopsy markers have been investigated in GCT, including cell-free nucleic acids like microRNAs, and circulating tumor cells (CTCs).Prognostic relevance has been demonstrated for circulating tumor cells (CTCs) in patients with different cancers. However, little is known in GCT patients. Histologically, GCT are a very heterogeneous group of tumors comprising pure seminomas (consisting of cells that remember primordial germ cells) and nonseminomas, which are either undifferentiated (embryonal carcinoma) or differentiated, exhibiting different degrees of embryonic (teratoma) or extraembryonic (yolk sac tumor and choriocarcinoma) differentiation. This heterogeneity hampers capture and detection of CTCs deriving from those tumors using a single method or a single antibody. To date, label-independent capture methods that enrich tumor cells according to the density of GCT cells, which is similar to that of mononuclear cells, have been successfully applied. Since testicular GCT might also express epithelial proteins, methods based on enrichment of CTCs using epithelial markers are promising to detect CTCs in certain subgroups of patients with GCTs as well.Here, we describe and discuss a combination of methods to capture and detect GCT cells with epithelial and germ cell characteristics in blood.


Assuntos
Biomarcadores Tumorais , Neoplasias Embrionárias de Células Germinativas/diagnóstico , Neoplasias Embrionárias de Células Germinativas/etiologia , Células Neoplásicas Circulantes/patologia , Neoplasias Testiculares/diagnóstico , Neoplasias Testiculares/etiologia , Linhagem Celular Tumoral , Separação Celular/métodos , Células Cultivadas , Imunofluorescência , Humanos , Imuno-Histoquímica , Hibridização in Situ Fluorescente , Biópsia Líquida , Masculino , Técnicas de Diagnóstico Molecular , Células Neoplásicas Circulantes/metabolismo , Prognóstico
17.
Cesk Patol ; 56(3): 153-160, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33076667

RESUMO

The great majority of testicular tumors can be diagnosed on the basis of morphology, while immunohistochemistry and molecular genetics assist in only a small proportion of cases. Similar to other areas of pathology, ancillary diagnostic methods have to be used responsibly and assessed in correlation with morphological, serological and clinical findings. Prior to their effective use, a limited differential diagnosis based on morphology is required.The significance of germ cell tumors is underscored by the fact that they represent the most frequent solid neoplasms occurring in men between 20-30 years and if diagnosed correctly and in early stage, they have excellent prognosis. From the molecular genetic standpoint, germ cell tumors stand apart from the current trend of tumor stratification based on molecular profiles. It is mainly due to the low mutational load, since the main genetic abnormality are chromosomal aneuploidies. Given the frequency of germ cell tumors among testicular neoplasms and since morphology is usually diagnostically most valuable, this review article is focused mainly on germ cell tumors, emphasizing the morphological features. Sertoli cell tumor, NOS is the only sex-cord stromal tumor included in this review as its diagnosis can be challenging. For practical purposes, this reviewis focused on differential diagnosis, including only entities where misdiagnosis would have impact on clinical outcome.


Assuntos
Neoplasias Embrionárias de Células Germinativas , Tumor de Células de Sertoli , Tumores do Estroma Gonadal e dos Cordões Sexuais , Neoplasias Testiculares , Biomarcadores Tumorais , Humanos , Imuno-Histoquímica , Masculino , Neoplasias Embrionárias de Células Germinativas/diagnóstico , Neoplasias Embrionárias de Células Germinativas/genética , Neoplasias Testiculares/diagnóstico , Neoplasias Testiculares/genética
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