The Capacity for Differentiation and Production of Human Chorionic Gonadotropin and α-Fetoprotein in Human Embryonal Carcinoma Cell Lines

Sekiya, Souei; Ishige, Hideo; Yamazawa, Koji; Kawata, Makoto; Inaba, Noriyuki; Nagao, Keiji; Takamizawa, H

doi:10.1097/00004347-198812000-00008

Cited by 9 publications

(9 citation statements)

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“…At 40 days after inoculation, tumor development was inspected. The tumors produced by inoculation of NEC14 cells into the intraabdominal cavity showed properties typical of teratocarcinoma consisting of differentiated elements such as cartilage, mesenchyme, gland and EC stem cells [29]. As previously shown [29], NEC14 cells were tumorigenic to nude mice when the cells were inoculated at a high dose (> lo').…”

Section: Establishment Of the Neci4 Cell Lines Expressing The N-myc Gmentioning

confidence: 62%

A transient decrease in N-myc expression and its biological role during differentiation of human embryonal carcinoma cells

Hasegawa¹,

Hara²,

Takehana³

et al. 1991

Differentiation

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Section: Establishment Of the Neci4 Cell Lines Expressing The N-myc Gmentioning

confidence: 62%

A transient decrease in N-myc expression and its biological role during differentiation of human embryonal carcinoma cells

Hasegawa¹,

Hara²,

Takehana³

et al. 1991

Differentiation

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“…Retinoic acid induced neural differentiation in the Tera 2 cells. Another human EC cell line, NEC 14, retained a pluripotent potential similar to that of murine EC cells and differentiated to form teratoma, choriocarcinoma or yolk sac elements [23]. Motoyama et al [24] showed that several human EC cell lines could differentiate into a YST in vitro and postulated that AFP production in EC is a functional expression of yolk sac differentiation [25].…”

Section: Discussionmentioning

confidence: 99%

Establishment of a testicular carcinoma cell line producing α‐fetoprotein

et al. 2001

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Objective To characterize a newly established human testicular carcinoma cell line that continuously produces a-fetoprotein (AFP), and to investigate the effects of retinoic acid on AFP production. Materials and methods A 24-year-old man underwent a radical orchidectomy for a right testicular tumour and was found to have two separate metastatic lesions in the lungs, both of which were removed surgically. The cancer cells were isolated from one of the tumours, which was composed of undifferentiated germ cells and produced AFP; the cells were cultured in a monolayer. This cell line was designated as KU-MT. Results The cell line was successfully maintained both in athymic nude mice and in culture. Histological examination showed that the xenografted tumours were composed of cells in the reticular, solid and glandular patterns of a yolk sac tumour, and of embryonal carcinoma cells. These cells immunostained positively for AFP. On electron microscopy, the extracellular deposition of a basement lamina-like substance, a typical feature of yolk sac tumour, was detected.The AFP production in mice correlated well with the tumour weight of the xenograft. The cultured KU-MT cells were oval to polygonal in morphology and grew exponentially, with a population doubling time of < 2 days. Chromosomal analysis showed a modal number of 57 with consistent structural abnormalities of +add(1)(p13), del(1)(q32), del(2)(q31), add(6)(q21), +add(9)(p22), add(11)(p15), and add(14)(p11). Reverse-transcription polymerase chain reaction analysis showed that the retinoic acid receptors (RAR)-a, RAR-c, and retinoid X receptor-a were present in the cells. The expression of AFP mRNA was up-regulated in response to all-trans-retinoic acid; treatment with this agent caused morphological changes and induced apoptosis in the cells. Conclusions This newly established cell line provides a reproducible model system that should offer a good insight into the differentiation of testicular carcinoma.

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“…The human EC cell line NEC14, established from a testicular germ cell tumor, has properties typical of EC cells (46,47). The stem cells are small and polygonal and form densely packed clusters that do not express a detectable level of FN on their surfaces.…”

Section: Discussionmentioning

confidence: 99%

“…The human EC cell line NEC14, derived from testicular germ cell tumors (47), was cultivated in RPMI 1640 medium supplemented with 10% fetal calf serum. The medium was changed every day, and the cells were subcultivated by repeated pipetting at a 1:3 to 1:4 dilution when they had reached confluence.…”

Section: Methodsmentioning

confidence: 99%

“…Owing to the shortage of established human cell lines that change their growth properties in response to external signals, the positive and negative transcription factors interacting with the G-rich sequences in the human FN promoter and its relation to the growth potential of cells have not yet been clarified. The human embryonal carcinoma (EC) cell line NEC14, established from a testicular germ cell tumor (46,47), offers a good system for analysis of these factors, since it proliferates unlimitedly but can be induced to differentiate by the addition of 10 Ϫ2 M N, NЈ-hexamethylene bisacetamide (HMBA) in vitro (20,48). The cells tend to lose proliferative potential upon induction of differentiation, appearing larger and flattened, and finally cease their growth.…”

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confidence: 99%

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Induction of Sp1 in Differentiating Human Embryonal Carcinoma Cells Triggers Transcription of the Fibronectin Gene

Suzuki

Oda

Nakajima

et al. 1998

Molecular and Cellular Biology

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Cells of the human embryonal carcinoma line NEC14 proliferate as densely packed clusters consisting of small, polygonal stem cells and do not express a detectable level of fibronectin (FN). Upon induction of differentiation by treatment with N,N-hexamethylene bisacetamide (HMBA), the level of FN mRNA increased steeply within 24 h and FN began to be accumulated, along with the organization of actin filaments in the cells. The FN promoter elements required for the activation were analyzed in reference to a cluster of GC boxes by using the chloramphenicol acetyltransferase (CAT) gene fused to 5 sequential-deletion derivatives of the promoter and promoters carrying base substitutions in the GC boxes. Among four GC boxes, GC boxes 2 and 3 had the greatest effect on promoter activation, and base substitutions in these GC boxes resulted in 80% reduction in promoter activity. The pattern of DNA-protein complex formation with these GC boxes changed drastically after induction of differentiation. The extract prepared from undifferentiated NEC14 cells formed fast-migrating complexes (UnD complexes), while the extract prepared from NEC14 cells treated with HMBA for 24 h formed slow-migrating complexes containing Sp1. Both complexes were formed predominantly with GC box 2. Base substitutions within the GC boxes completely abolished the formation of both UnD and Sp1 complexes. Consistent with these changes, the Sp1 level increased steeply within 24 h. Induction of Sp1 expression in NEC14 cells effectively stimulated the promoter activity of the transfected FN promoter-CAT constructs. These results indicate that activation of the FN promoter in differentiating NEC14 cells occurs by the steep induction of Sp1, which prevents an undifferentiated cell factor from binding to the Sp1 sites.Fibronectin (FN) is a family of large glycoproteins of the extracellular matrix and has multiple domains with specific binding sites for other matrix macromolecules, such as fibrin, glycosaminoglycans, and collagen, and for its receptor, the ␣5␤1-type integrin on the cell surface (24). FN binds to its receptor as a dimer of similar subunits of about 250 kDa (23,24). The cytoplasmic domain of the receptor interacts with actin filaments via several adhesion proteins, connecting the extracellular matrix to actin filaments at the receptor site called focal contacts or adhesion plaques (3,4,23,25,44). FN thus plays an important role in organizing the extracellular matrix and enabling cells to attach to it. Through these functions, FN regulates cell adhesion, migration, growth, wound healing, and tumor metastasis (12,23).Expression levels of FN in cells vary markedly depending on the environment and cellular capacity for proliferation. Cells transformed by various oncogenes usually express very low levels of FN (14,17,29), while senescent cells inevitably express high levels of FN, ceasing their proliferation (18,35,39). We previously showed that the level of FN gene expression in the rat derivative cell line 3Y1 transformed by adenovirus E1A and E1B g...

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The Capacity for Differentiation and Production of Human Chorionic Gonadotropin and α-Fetoprotein in Human Embryonal Carcinoma Cell Lines

Cited by 9 publications

References 0 publications

A transient decrease in N-myc expression and its biological role during differentiation of human embryonal carcinoma cells

A transient decrease in N-myc expression and its biological role during differentiation of human embryonal carcinoma cells

Establishment of a testicular carcinoma cell line producing α‐fetoprotein

Induction of Sp1 in Differentiating Human Embryonal Carcinoma Cells Triggers Transcription of the Fibronectin Gene

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