Isolation of Hormone-Producing Clonal Lines of Human Choriocarcinoma<sup>1</sup>

Kohler, Peter O.; Bridson, William E.

doi:10.1210/jcem-32-5-683

Cited by 356 publications

(141 citation statements)

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“…JEG-3 choriocarcinoma cells [30] were grown in DMEM supplemented with 10% FCS, penicillin (100 U/ml) and streptomycin (100 μg/ml). HIT cells were transfected by the diethylaminoethyl (DEAE)-dextran method [31] with 2 μg of the luciferase reporter gene and the expression vector as indicated.…”

Section: Plasmid Constructionmentioning

confidence: 99%

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Inhibition of MafA transcriptional activity and human insulin gene transcription by interleukin-1β and mitogen-activated protein kinase kinase kinase in pancreatic islet beta cells

et al. 2007

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Aims/hypothesis Inappropriate insulin secretion and biosynthesis are hallmarks of beta cell dysfunction and contribute to the progression from a prediabetic state to overt diabetes mellitus. During the prediabetic state, beta cells are exposed to elevated levels of proinflammatory cytokines. In the present study the effect of these cytokines and mitogen-activated protein kinase kinase kinase 1 (MEKK1), which is known to be activated by these cytokines, on human insulin gene (INS) transcription was investigated. Methods Biochemical methods and reporter gene assays were used in a beta cell line and in primary pancreatic islets from transgenic mice. Results IL-1β and MEKK1 specifically inhibited basal and membrane depolarisation and cAMP-induced INS transcription in the beta cell line. Also, in primary islets of reporter gene mice, IL-1β reduced glucose-stimulated INS transcription. A 5′-and 3′-deletion and internal mutation analysis revealed the rat insulin promoter element 3b (RIPE3b) to be a decisive MEKK1-responsive element of the INS. RIPE3b conferred strong transcriptional activity to a heterologous promoter, and this activity was markedly inhibited by MEKK1 and IL-1β. RIPE3b is also known to recruit the transcription factor MafA. We found here that MafA transcription activity is markedly inhibited by MEKK1 and IL-1β. Conclusions/interpretation These data suggest that IL-1β through MEKK1 inhibits INS transcription and does so, at least in part, by decreasing MafA transcriptional activity at the RIPE3b control element. Since inappropriate insulin biosynthesis contributes to beta cell dysfunction, inhibition of MEKK1 might decelerate or prevent progression from a prediabetic state to diabetes mellitus.

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Section: Plasmid Constructionmentioning

confidence: 99%

“…Islets were collected, washed once with PBS and resuspended in potassium phosphate buffer, pH 7.8, followed by three freeze-thaw cycles. Luciferase activity [30] and protein (Protein Assay; Bio-Rad Laboratories, Munich, Germany) content were determined in the supernatant.…”

Section: Plasmid Constructionmentioning

confidence: 99%

Inhibition of MafA transcriptional activity and human insulin gene transcription by interleukin-1β and mitogen-activated protein kinase kinase kinase in pancreatic islet beta cells

et al. 2007

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“…Among these are naturally immortalized cell lines obtained from choriocarcinoma tissue, such as BeWo [16], JAR [17] and JEG-3 [18]. The cell lines AC1M-32 [19] and ACH-3P [20] have been generated by fusion of the AC1-1 cell line (a JEG-3 mutant [21]) with primary term or first trimester trophoblasts, respectively.…”

Section: Introductionmentioning

confidence: 99%

Toll-like receptor profiling of seven trophoblast cell lines warrants caution for translation to primary trophoblasts

et al. 2015

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“…In samples from first trimester miscarriage, DNA and proteins of AAV were detected predominantly in trophoblast cells (Tobiasch et al, 1994). This finding prompted us to analyse the choriocarcinoma-derived human trophoblast cell lines BeWo (Pattillo & Gey, 1968), JEG-3 (Kohler & Bridson, 1971) and JAr (Pattillo et al, 1971), as well as the human amnion-derived FL cell line (Fogh & Lund, 1957), for AAV. Analysis by PCR using primers amplifying sequences of AAV (Tobiasch et al, 1994) indicated the presence of DNA of AAV type 2 (AAV-2) within the human trophoblast cell lines (Fig.…”

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confidence: 99%

Presence of integrated DNA sequences of adeno-associated virus type 2 in four cell lines of human embryonic origin.

Dutheil

Malhomme

Provost

et al. 1997

Journal of General Virology

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The human helper virus-dependent parvovirus adeno-associated virus (AAV) has been found in human female genital tissues including material from first trimester miscarriage. In the latter case, AAV type 2 (AAV-2) DNA and viral proteins were detected mainly in the trophoblast cell layer of placenta. In this report, we present evidence that AAV DNA is also present in established human trophoblast cell lines (JEG-3, JAr, BeWo) and in the human amnion cell line FL. In cells of these lines, AAV-2 DNA could be detected both by PCR and Southern blot analysis. Restriction enzyme analysis indicated that AAV DNA was integrated into the host cell genome. Although the cell lines supported AAV replication when infected with AAV-2 and adenovirus type 2 (Ad2) as a helper virus, superinfection with Ad2 alone did not induce replication of AAV DNA, i.e. it failed to rescue AAV from its integrated state. This is probably due to rearrangements within the integrated AAV genome.

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Isolation of Hormone-Producing Clonal Lines of Human Choriocarcinoma¹

Cited by 356 publications

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Inhibition of MafA transcriptional activity and human insulin gene transcription by interleukin-1β and mitogen-activated protein kinase kinase kinase in pancreatic islet beta cells

Inhibition of MafA transcriptional activity and human insulin gene transcription by interleukin-1β and mitogen-activated protein kinase kinase kinase in pancreatic islet beta cells

Toll-like receptor profiling of seven trophoblast cell lines warrants caution for translation to primary trophoblasts

Presence of integrated DNA sequences of adeno-associated virus type 2 in four cell lines of human embryonic origin.

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Isolation of Hormone-Producing Clonal Lines of Human Choriocarcinoma1

Cited by 356 publications

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Inhibition of MafA transcriptional activity and human insulin gene transcription by interleukin-1β and mitogen-activated protein kinase kinase kinase in pancreatic islet beta cells

Inhibition of MafA transcriptional activity and human insulin gene transcription by interleukin-1β and mitogen-activated protein kinase kinase kinase in pancreatic islet beta cells

Toll-like receptor profiling of seven trophoblast cell lines warrants caution for translation to primary trophoblasts

Presence of integrated DNA sequences of adeno-associated virus type 2 in four cell lines of human embryonic origin.

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Isolation of Hormone-Producing Clonal Lines of Human Choriocarcinoma¹