bromochloroacetic-acid and Seminoma

To describe the establishment and characterization of a human cell line, SEM-1, from a patient diagnosed with a mediastinal seminoma.. A small percentage of germ cell tumors develop as primary lesions in extragonadal sites, and the etiology of these tumors is poorly understood. Currently, only 2 cell lines from seminoma patients have been reported, JKT-1 and TCam-2, both derived from the testis. The cell line was characterized by heterotransplantation in Nude mice, cytogenetic studies, immunohistochemical and flow cytometry staining for germ cell tumor biomarkers, quantitative reverse-transcription polymerase chain reaction for cancer testis antigen expression, and BRAF mutation screening with quantitative polymerase chain reaction.. Characterization studies confirmed the human extragonadal seminoma origin of SEM-1 and demonstrated that it had more features in common with TCam-2 than JKT-1. Specifically, SEM-1 was positive for Sal-like protein 4 (SALL-4), activator protein-2γ (AP-2γ), and cytokeratin CAM5.2, and demonstrated heterogeneous expression of stem cell markers octamer-binding transcription factor 3/4, NANOG, c-KIT, SOX17, and SOX2. Cytogenetic analysis revealed a hypotriploid chromosome number, with multiple copies of 12p, but isochromosome 12p and the BRAF mutation V600E were not identified. The cell lines also did not contain the BRD4/NUT gene rearrangement [t(15,19)] seen in midline carcinomas nor did they contain overexpressed nuclear protein in testis (NUT) genes.. SEM-1 is the first cell line derived from an extragonadal germ cell tumor showing intermediate characteristics between seminoma and nonseminoma, and as such, is an important model to study the molecular pathogenesis of this malignancy.

Topics: Aneuploidy; Biomarkers; Biomarkers, Tumor; Cell Line, Tumor; DNA Mutational Analysis; Gene Expression; Gene Rearrangement; Homeodomain Proteins; Humans; Keratins; Male; Middle Aged; Nanog Homeobox Protein; Neoplasm Proteins; Nuclear Proteins; Octamer Transcription Factor-3; Oncogene Proteins; Proto-Oncogene Proteins B-raf; Proto-Oncogene Proteins c-kit; RNA, Messenger; Seminoma; SOXB1 Transcription Factors; SOXF Transcription Factors; Transcription Factor AP-2; Transcription Factors

Topics: Adult; Antigens, CD20; Carcinoma; Diagnosis, Differential; DNA Nucleotidylexotransferase; Female; Follow-Up Studies; Humans; Keratins; Lymphoma; Male; Middle Aged; Necrosis; Seminoma; Thymoma; Thymus Neoplasms; Tuberculosis

Necrotic testicular tumors are relatively frequent and can present a significant diagnostic challenge. Because of differing treatments for seminomas versus nonseminomas, accurate diagnosis is critical. Eleven totally (n=9) or almost totally (n=2) necrotic testicular tumors were retrieved from our consult files. The submitting pathologists favored benign processes in 4 cases, Leydig cell tumor in 1, and lymphoma in 1. The cases were evaluated for histologic features and, when material was available, by immunostaining with 7 antibodies: keratin (AE1/AE3), OCT4, placental alkaline phosphatase, alpha-fetoprotein (AFP), CD117, CD30, and S100. Only distinct reactivity in a cellular distribution in the necrotic zone was considered positive; nuclear reactivity alone was scored for OCT4 and membrane reactivity for CD117 and CD30. Mean patient age was 35 years (range 16-63). Mean tumor size was 19 mm (range 7-53). All patients presented with unilateral testicular masses (6 right, 5 left); 2 also had acute pain. The combination of histologic features, immunostains and, in 1 case, serum AFP permitted classification of 8 tumors (4 seminomas, 3 embryonal carcinomas, 1 yolk sac tumor). Three were not classifiable. The necrotic seminomas lacked associated coarse intratubular calcifications and were positive for OCT4 (4/4) and CD117 (3/3) but negative for keratin (0/4) and CD30 (0/4). The necrotic embryonal carcinomas had associated coarse intratubular calcifications and were positive for keratin (2/3), OCT4 (2/2), and CD30 (3/3). OCT4 stained 1 unclassifiable tumor, which lacked other specific markers. We did not find placental alkaline phosphatase, AFP, and S100 stains useful, although S100 did highlight tumor "ghost" cells in 1 case. Other features in most cases included intratubular germ cell neoplasia (6/11), tubular atrophy/hyalinization (10/11), tumor "ghost" cells (10/11), scar (9/11), and inflammation (10/11). Of the 5 patients with available follow-up, 3 were free of disease at 1, 5, and 8 years after orchiectomy (2 necrotic seminomas and 1 germ cell tumor, unclassified). One patient with yolk sac tumor (age 63 y) developed widespread metastases after 15 months and died of disease. The final case was initially misinterpreted as "testicular infarction, no malignancy" and 16 months later the patient developed a large retroperitoneal seminoma. Most totally necrotic testicular tumors can be placed into clinically important groups by assessment for coarse intratubula

Topics: Adolescent; Adult; alpha-Fetoproteins; Biomarkers, Tumor; Carcinoma, Embryonal; Disease-Free Survival; Endodermal Sinus Tumor; Humans; Immunohistochemistry; Keratins; Ki-1 Antigen; Male; Middle Aged; Necrosis; Octamer Transcription Factor-3; Orchiectomy; Proto-Oncogene Proteins c-kit; Seminoma; Testicular Neoplasms; Young Adult

In spite of the high curability rates, rare cases of testicular seminoma behave in an unexpectedly aggressive manner. No effective markers are currently available that can predict such uncommon behavior. We studied 105 cases of testicular seminoma on whom the primary resection was performed at the Memorial Sloan-Kettering Cancer Center, New York, to investigate any relationship between morphology, immunohistochemical features, and clinical/pathological stages. Fifty-nine percent of the cases presented with pT stage 1 and 74 percent with American Joint Committee on Cancer (AJCC) stage I. In univariate analysis, tumor size, mitotic count, and presence of necrosis showed significant associations with pT stage (p = 0.0001, 0.0001, and 0.04, respectively), and the presence of vascular invasion (p = 0.0001, 0.0001, and 0.02, respectively). In multivariate analysis, both the tumor size and mitotic counts were independent predictors of pT stage (p = 0.0004 and 0.0001) and vascular invasion (p = 0.0004 and 0.0001). When tumors were separated on the basis of architectural and/or cytological atypia into "usual seminomas" and "seminomas with atypia", these were significantly associated with AJCC stage (p = 0.02), and c-kit protein (p = 0.0005) and CD30 expression (p = 0.02). In addition, "seminomas with atypia" also tended to show a higher proliferation activity as judged by Ki67 reactivity (p = 0.06), as well as express the marker of epithelial differentiation, Cam 5.2 (p = 0.09). In summary, we find that the morphologic features in testicular seminomas are associated with factors of clinical relevance. Also, "seminomas with atypia" differ from "usual seminomas" morphologically, present at a higher AJCC stage, and possibly represent an early step in transformation of seminomas toward a more aggressive phenotype. While not proposing a new entity, we suggest that when these atypical features are encountered in an otherwise classical seminoma, investigations must be performed to exclude an early carcinomatous differentiation or even earlier changes toward such a differentiation, such as lack of c-kit protein expression.

Topics: Adult; Biomarkers; Biomarkers, Tumor; Humans; Immunohistochemistry; Keratins; Ki-1 Antigen; Ki-67 Antigen; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Staging; Prognosis; Proto-Oncogene Proteins c-kit; Seminoma; Testicular Neoplasms

The distinction of Sertoli cell tumors from seminoma is critical to ensure proper treatment. Although usually straightforward, we highlight herein 13 malignant Sertoli cell tumors of the testis with light microscopic features that mimicked seminoma. All of the cases were received in consultation, 10 with submitting diagnoses of seminoma, usually of classic type, but three cases of spermatocytic type. Patients ranged from 15 to 80 years of age (median 37 years); all presented with testicular masses. The tumors were typically firm, white to yellow-tan, and often had foci of hemorrhage. The dominant microscopic pattern was nested or sheet-like, with some tumors having secondary patterns of trabeculae-solid tubules, hollow tubules, and pseudofollicles. Tumor cells were polygonal with conspicuous clear cytoplasm in 12 cases; the cytoplasm was focally eosinophilic in 10 cases, but this was never conspicuous. Nine tumors had cytoplasmic vacuoles, and three of four that were investigated stained for intracytoplasmic glycogen. Nuclei were small (5) to medium-sized (8), round-to-oval (13), and vesicular with irregular contours (11). Nucleoli were present in 11 tumors (six small; five large). Stromal fibrosis (12) and lymphoid infiltrates (10) were conspicuous, and tumor necrosis (11) and vascular invasion (8) also were seen. Mitotic figures ranged from <1 to 21/10 high power fields (HPF) (median 1/10 HPF). Staining for inhibin-alpha, epithelial membrane antigen, and cytokeratin (AE1/AE3) was positive in four of four, six of six, and three of six cases, respectively; placental alkaline phosphatase was negative in all five tumors investigated. The nested growth pattern, prominence of clear cells, lymphoid infiltrate, inconspicuous tubular differentiation, cytoplasmic glycogen, and prominent nucleoli caused these tumors to be mistaken for seminomas. The smaller, less pleomorphic nuclei of Sertoli cell tumors, their lower mitotic rate, and the absence of intratubular germ cell neoplasia are helpful differential features. Immunohistochemistry is a useful adjunct in confirming the diagnosis of Sertoli cell tumor, but only if the overlapping features are appreciated by conventional microscopy and the diagnosis of Sertoli cell tumor included in the differential.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Diagnosis, Differential; Humans; Immunoenzyme Techniques; Inhibins; Keratins; Male; Middle Aged; Mucin-1; Neoplasm Proteins; Seminoma; Sertoli Cell Tumor; Testicular Neoplasms

We studied cytokeratin (CK) expression immunohistochemically in 64 seminomas using a panel of commercially available antikeratin antibodies and tested for association of CK expression with patient age, tumor size, stage, and outcome. Seventeen embryonal carcinomas were compared with seminoma. CK7, CAM 5.2, AEI/AEIII, and wide-spectrum screening keratin (WSK) were positive in 41%, 30%, 36%, and 36% of the seminomas, respectively. CK20 and high-molecular-weight keratin (HMWK) were negative in all cases. CD30, placental alkaline phosphatase (PLAP), and epithelial membrane antigen (EMA) were positive in 6%, 100%, and 2% of cases, respectively. There were no differences in patient age, stage, tumor size, or outcome between CK-positive and CK-negative seminomas. CK7, CAM 5.2, AEI/AEIII, and WSK were positive in 100%, 88%, 94%, and 88% of embryonal carcinomas, respectively. CK20 and HMWK were negative in all cases. CD30, EMA, and PLAP were positive in 100%, 12%, and 76%, respectively. CKs are present in seminoma, and their presence is not associated with a difference in patient age, stage, or outcome. In cases such as small needle biopsy specimens, CK and CD30 stains may be useful in separating seminoma from embryonal carcinoma.

Topics: Adult; Aged; Alkaline Phosphatase; Carcinoma, Embryonal; Germinoma; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Mucin-1; Neoplasm Staging; Seminoma; Testicular Neoplasms

Based on immunohistochemistry (IHC) and DNA ploidy, different paths of carcinogenesis have been suggested for spermatocytic seminoma (SS) and classical seminoma (CS). The present study extends current knowledge on the above parameters.. Seventeen SSs and twenty-two CSs were assessed by IHC for placental-like alkaline phosphatase (PLAP), c-kit, cytokeratin and adhesion carbohydrate molecyles. All SSs and 11 CSs were also analysed for DNA ploidy.. All CSs, but none of the SSs, were positive for PLAP. C-kit positivity was found in 7 of 17 SSs and in all CSs. The other IHC parameters were similarly distributed among the evaluated SSs and CSs. Fourteen SSs were diploid or polyploid, and three were aneuploid. All CSs were aneuploid.. The new observation of c-kit positivity in about 40% of SSs suggests that at least some of the SSs originate from primordial cells. The predominantly diploid or polyploid DNA pattern indicates that SSs follow a pathogenetic pathway which is most probably different from that of CSs.

Topics: Adult; Aged; Aged, 80 and over; Alkaline Phosphatase; Aneuploidy; Cell Adhesion Molecules; Diploidy; DNA, Neoplasm; Flow Cytometry; Humans; Immunohistochemistry; Isoenzymes; Keratins; Male; Middle Aged; Ploidies; Polyploidy; Proto-Oncogene Proteins c-kit; Seminoma; Testicular Neoplasms

A case of tubular seminoma in a 33-year-old man is reported. The tumor occurred in his right testis, measuring 7 x 6 x 6 cm in size. Microscopically, most of the tumor cells were arranged in solid or hollow tubular patterns. The tumor cells had round nuclei with vesicular chromatin and clear or granular cytoplasm, characteristic features of seminoma cells. A classic seminoma component and intratubular atypical germ cells occupying approximately 5% of the tumor area were noted. Immunohistochemical staining revealed that the tumor cells were diffusely placental alkaline phosphatase and vimentin positive, and focally cytokeratin positive. These findings are consistent with a tubular seminoma. The differential diagnoses are discussed.

Topics: Adult; Alkaline Phosphatase; GPI-Linked Proteins; Humans; Immunohistochemistry; Isoenzymes; Keratins; Male; Seminoma; Testicular Neoplasms; Treatment Outcome; Vimentin

Topics: Humans; Keratins; Seminoma

In testicular germ cell tumors the CD30 antigen has been shown to be regularly expressed in embryonal carcinoma and was thus suggested as a marker for this particular neoplasm. Very recently, it has been proven that the monoclonal antibody Ber-H2 is suitable for the detection of this membrane antigen in paraffin sections. We conducted an immunohistochemical study to investigate the CD30 expression in a large series of different presentations of seminoma (ie, pure, mixed, and spermatocytic) because there is evidence from several sources that embryonal carcinoma is histogenetically closely related to, and probably derives from, seminoma. Sections from formalin-fixed, paraffin-embedded tissue from 38 cases of testicular seminomas were immunostained for the demonstration of the CD30 antigen using the monoclonal antibody Ber-H2, cytokeratins, and placental alkaline phosphatase following an indirect streptavidin-peroxidase regimen. In selected cases, immunostainings were performed on consecutive sections to investigate a possible colocalization of CD30 and cytokeratins in seminoma. Specific immunostaining for CD30 in seminoma cells could be detected in single minute foci in 4 of 21 cases of pure classic seminoma. Seminomatous components of mixed tumors showed CD30 positivity in single, but also multiple, foci in 7 of 14 cases. CD30 immunoreactivity in seminoma cells occurred with and without colocalized expression of cytokeratin. Spermatocytic seminoma (n = 3) as well as intratubular germ cell neoplasia in tumor adjacent parenchyma (n = 36) were negative in all cases investigated. We conclude that in testicular germ cell tumors, the expression of CD30 is not restricted to embryonal carcinoma but can also be found focally in seminoma, adding further evidence for a close relationship between these two tumors. The prevalence of CD30 expression in seminomatous components of mixed tumors, as well as the coexpression with cytokeratins, suggest that CD30 expression in seminomas might indicate their upcoming transformation to embryonal carcinoma. This conclusion coincides with a model featuring seminoma in a central role of germ cell tumor development.

Topics: Adolescent; Adult; Aged; Alkaline Phosphatase; Biomarkers, Tumor; Carcinoma, Embryonal; Humans; Immunohistochemistry; Keratins; Ki-1 Antigen; Male; Middle Aged; Seminoma; Testicular Neoplasms

Intratubular germ cell neoplasia (ITGCN) is now considered to be the preinvasive phase of testicular germ cell tumors with the exceptions of spermatocytic seminoma, pure yolk sac tumor, and mature teratoma. Pagetoid spread of ITGGN into rete testis is a common yet unpublished finding in these cases. We reviewed 100 cases of testicular germ cell tumors from the Surgical Pathology service of Parkland Memorial Hospital (Dallas, TX) to evaluate the frequency of this pattern of spread. Additional sections were obtained from selected cases and were stained with anti-placental alkaline phosphatase, anti-low molecular weight keratin (clone AE1), and various lectins to highlight the process. Pagetoid spread of ITGCN into rete testis was identified in 24 of 60 cases (40%) in which histologic sections contained both ITGCN and rete testis. The incidence of pagetoid ITGCN involvement of the rete testis was lower in pure seminoma (seven of 25 cases [28%]) than in testes containing nonseminomatous germ cell tumors (17 of 35 cases [49%]). AE1 stained the epithelial cells of the rete testis but not the cells of the ITGCN, whereas placental alkaline phosphatase stained the neoplastic cells but not the epithelial cells of the rete testis. These stains were useful in delineating two cases in which the pagetoid involvement was so extensive that they were misdiagnosed as invasive seminomas. Pagetoid spread of ITGCN is a relatively common finding in testicular germ cell tumors and rarely can be mistaken for invasive seminoma. Immunohistochemistry can be helpful in distinguishing florid pagetoid spread from invasive seminoma.

Topics: Adolescent; Adult; Alkaline Phosphatase; Carcinoma in Situ; Cell Transformation, Neoplastic; Diagnosis, Differential; Epithelium; Germinoma; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Orchiectomy; Paget Disease, Extramammary; Seminoma; Testicular Neoplasms