Establishment and Characterization of a New Human Testicular Seminoma Cell Line, JKT‐1

Kinugawa, Keigo; Hyodo, Fuminori; Matsuki, Takakazu; Jo, Yoshimasa; Furukawa, Yoji; Ueki, Ayako; Tanaka, Hiroyoshi

doi:10.1111/j.1442-2042.1998.tb00604.x

Cited by 41 publications

(54 citation statements)

References 10 publications

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“…This technique was modified and simplified several times and recently it has been widely used for the initiation of primary cultures of adherent cells (Li et al 2009;Wang et al 2003;Kinugawa et al 1998).…”

Section: Resultsmentioning

confidence: 99%

Biological and morphological characterization of human neonatal fibroblast cell culture B-HNF-1

et al. 2010

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In the present study, human neonatal fibroblasts were isolated from a two-month-old human male. The purpose of the present investigation was the analysis of the morphology (light and transmission electron microscopy), karyotype and growth characteristics of the human neonatal fibroblast cell culture B-HNF-1. Moreover, STR typing and mitochondrial DNA amplification and sequencing was also performed. Analysis of chromosomes count showed that B-HNF-1 cell culture is diploid and has normal male karyotype 46, XY, which was stable during cultivation. The transmission electron microscopy demonstrated the ultra-structure of the B-HNF-1 cells; they have typical morphological features of proteosynthesis-active cells. Large number of fibroblasts bearing different shapes and surface characteristics adhered to the substrate with microvilli and filopodia. Our in vitro expanded fibroblasts have a large and irregular nucleus with prevalence of euchromatin. One to three nucleoli are present in each nucleus. The cytoplasm contains a richly developed rough endoplasmic reticulum and prominent Golgi apparatus cisterns. The result of the fragment analysis is a DNA profile defined as multiplex of STR markers and sex determining system. Sequencing analysis of hypervariable segments of control region of mitochondrial DNA showed haplotype that belongs to haplogroup W. In this study, we complexly characterized a new cell culture of human neonatal fibroblasts B-HNF-1 from different points of view. This culture should be used for further biomedical experiments.

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Section: Resultsmentioning

confidence: 99%

Biological and morphological characterization of human neonatal fibroblast cell culture B-HNF-1

et al. 2010

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“…The three testicular tumors were classified as pure seminoma by histological analysis and by positive staining for placental alkaline phosphatase (PlAP), a specific seminoma marker (Giwercman et al 1991). The JKT-1 cells, described as a pure human seminoma cell line (Kinugawa et al 1998), also expressing PIAP (Roger et al 2004), were maintained in DMEM supplemented with 10% fetal bovine serum (FBS) (Gibco BRL). For estrogen stimulation, JKT-1 cells were plated in the above complete medium, estrogen-starved for 24 h in fresh phenol red-free DMEM supplemented with 10% charcoal-stripped serum (3 g charcoal-coated dextran/50 ml FBS), before adding every culture day 17 -estradiol (Sigma), ICI 182780 (Falsodex; Astra-Zeneca, Birmingham, UK) or ethanol as vehicle control.…”

Section: Tissue Preparation and Cell Culturementioning

confidence: 99%

“…However, little is known of the possible estrogen-dependency of seminoma, the most frequent malignant testicular germ cell proliferation, partly due to the lack of available in vitro models. We therefore took advantage of a pure human testicular seminoma cell line, JKT-1 (Kinugawa et al 1998, Roger et al 2004, to assess the in vitro effects of estrogens on seminoma cell proliferation, to characterize ER subtypes and to test for a functional aromatase complex.…”

Section: Introductionmentioning

confidence: 99%

Estrogen-induced growth inhibition of human seminoma cells expressing estrogen receptor β and aromatase

Roger¹,

Lambard²,

Bouskine³

et al. 2005

Journal of Molecular Endocrinology

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It is now well established that estrogens participate in the control of normal spermatogenesis and endogenous or environmental estrogens are involved in pathological germ cell proliferation including testicular germ cell tumors. Studying a human testicular seminoma cell line, JKT-1, we show here that 17 -estradiol (10 −12 to 10 −6 M) induced in vitro a significant dose-dependent decrease of cell growth. This antiproliferative effect was maximum after 4 days of exposure at a physiologically intratesticular concentration of 10 −9 M, close to the K d of ER, and reversed by ICI 182780, an ER antagonist, suggesting an ER-mediated pathway. By RT-PCR and Western blot we were able to confirm that JKT-1, like tumoral seminoma cells and normal human testicular basal germ cells, expresses estrogen receptor (ER ), including ER 1 and ER 2, a dominant negative variant, but not ER . Using immunofluorescence and confocal microscopy, ER was observed as perinuclear intracytoplasmic spots in JKT-1 and tumoral seminoma cells without significant translocation of ER into the nucleus, under 17 -estradiol exposure. Double staining observed by confocal microscopy revealed that ER colocalized in JKT-1 cells with cytochrome C, a mitochondrial marker. We report for the first time the expression of a functional aromatase complex in seminoma cells as assessed by RT-PCR, Western blot and enzymatic assay. Seminoma cells are able to respond to estrogens through a possible autocrine or paracrine loop. These preliminary results support estrogen-dependency of human testicular seminoma, the most frequent tumor of young men, and suggest potential pharmacological use. Whether this estrogen control, however, involves an ER -mediated stimulation of cell apoptosis and/or an ER -mediated inhibition of cell proliferation, remains to be further determined.

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“…However, with the exception of primary tumor samples, few tools are available to study the molecular pathogenesis of seminoma (15). To the best of our knowledge and according to the literature, only 3 cell lines (TCam-2, JKT-1 and SEM-1) originate from seminoma (16)(17)(18). Furthermore, the origin of these cell lines is not certain; therefore it remains controversial to describe these cell lines as seminoma cell lines (19,20).…”

Section: Discussionmentioning

confidence: 99%

In vitro study on shRNA-mediated reduction of testis developmental related gene 1 expression and its effects on the proliferation, invasion and apoptosis of NTERA-2 cells

et al. 2015

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Abstract. Testis developmental related gene 1 (TDRG1) is a novel human testis-specific gene. TDRG1 is differentially expressed in cancerous tissue compared with normal testicular tissue and demonstrates a unique expression pattern in normal testes; therefore, this gene may be involved in the occurrence and development of testicular germ cell tumors (TGCT). In the present study, the expression level of TDRG1 was downregulated in human TGCT NTERA-2 cells by RNA interference (RNAi) in order to investigate the association between TDRG1 and TGCT. The TDRG1 mRNA and protein expression levels in NTERA-2 cells were significantly inhibited following transfection with specific RNAi plasmids. The ability to proliferate (inhibited by 15.4% at day 3 and 26.1% at day 5; P<0.001) and invade (reduced by 49.1%; P=0.01) in vitro was suppressed in cells in which the expression level of TDRG1 was reduced, and a corresponding increase in the apoptotic potential was observed (the early apoptotic potential and total apoptotic potential were increased by 75%; P=0.019 and 54.8%; P=0.009, respectively). The results of the present study indicated that the biological behavior of NTERA-2 cells is associated with TDRG1 expression levels, and that this gene may be a novel target candidate in the treatment of TGCT.

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Establishment and Characterization of a New Human Testicular Seminoma Cell Line, JKT‐1

Abstract: This report profiled a seminoma cell line established for both in vitro and in vivo experimental systems. Future studies are planned to investigate germ cells using this seminoma line.

Cited by 41 publications

References 10 publications

Biological and morphological characterization of human neonatal fibroblast cell culture B-HNF-1

Biological and morphological characterization of human neonatal fibroblast cell culture B-HNF-1

Estrogen-induced growth inhibition of human seminoma cells expressing estrogen receptor β and aromatase

In vitro study on shRNA-mediated reduction of testis developmental related gene 1 expression and its effects on the proliferation, invasion and apoptosis of NTERA-2 cells

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