Human thyroid epithelial cells cultured in monolayers. I. Decreased thyroglobulin and cAMP response to TSH in 12-week-old secondary and tertiary cultures

Rasmussen, Åse Krogh; Kayser, Lars; Perrild, Hans; Brandt, M. R.; Bech, K; Feldt‐Rasmussen, Ulla

doi:10.1016/0303-7207(95)03711-x

Cited by 22 publications

(16 citation statements)

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“…Though HepG2 cells do not synthesize insulin, they are glucose-responsive and increase expression of genes encoding metabolic enzymes, such as fatty acid synthase, after glucose stimulation [34]. Stimulation of primary human thyrocyte cultures [22] with TSH also did not affect egr-1 mRNA expression (Fig. 2D), demonstrating that egr-1 induction is not a general feature of activation, increased intracellular cAMP or exocytosis in other endocrine cell types.…”

Section: Resultsmentioning

confidence: 99%

“…Primary rat liver cells, obtained by in situ collagenase digestion, were grown in defined medium with or without insulin [21]. Human thyroid cells were obtained as surgical specimens of healthy gland adjacent to non-toxic thyroid adenomas and seeded as monolayers in RPMI 1640 containing growth factors (insulin, somatostatin, transferrin, hydrocortisone and glycyl-histidyl-lysine acetate) [22]. After 48 h in vitro culture, cells were stimulated for 30 min with 1 U/l TSH.…”

Section: Methodsmentioning

confidence: 99%

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Glucose induces early growth response gene (Egr-1) expression in pancreatic beta cells

et al. 1999

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Glucose homeostasis is maintained by beta cells of pancreatic islets through precisely regulated release of insulin from preformed granules. To maintain constant intracellular insulin stores while meeting wide physiological variations in demand, insulin synthesis is modulated in parallel with secretion rates [1]. Immediately following glucose stimulation, synthesis is primarily augmented through enhanced translation [2] and decreased degradation of existing insulin mRNAs [3]. Glucose also induces delayed responses that are manifested several hours after stimulation [4] by increasing insulin gene transcription [5] and expression of other genes involved in beta-cell function. Increased gene expression of glucose transporter 2 (GLUT-2) [6], pyruvate kinase [7], acetyl-coenzyme A-carboxylase [8], and 64 kDa glutamic acid decarboxylase (GAD65) [9] have been observed in primary rat islets in vitro and in rodent insulin-producing tumour cell lines following exposure to glucose. Together these changes comprise a delayed adaptive response that could be necessary to meet increased metabolic and secretory demands during extended or repeated periods of hyperglycaemia. Diabetologia (1999) Summary A copy deoxyribonucleic acid (cDNA) clone of the immediate early growth response gene, egr-1 (Krox-24, Zif268, NGFI-1), was isolated through subtractive hybridization screening to identify glucose-induced genes in pancreatic beta cells. Glucose rapidly and transiently induced egr-1 mRNA in the SV40-transformed murine beta-cell line, MIN6. Glucose also increased egr-1 mRNA expression in INS-1, bTC3 and RINm5F beta-cell lines, although with different kinetics. Expression of the 82 kDa Egr-1 protein was induced both in MIN6 cells stimulated with glucose in vitro and in primary rat islet cells stimulated in vivo or in vitro. This response is unique to beta cells since glucose did not affect egr-1 expression in NIH-3T3 fibroblasts or glucosesensitive hepatocytes. In beta cells egr-1 induction is specifically associated with insulin secretion, as it was not observed after stimulation with serum or insulin but was elicited by insulin secretagogues, including membrane depolarizing agents and cAMP agonists. Moreover, induction of egr-1 by glucose was inhibited by EDTA, indicating dependence on influx of extracellular Ca 2+. Other immediate early response genes, c-fos and junB, were also induced following glucose stimulation with kinetics similar to egr-1, whereas c-jun and junD expression were not affected. Since the zinc-finger protein encoded by egr-1 is highly homologous to transcription factors that control expression of glucose-regulated genes in yeast, Egr-1 could mediate delayed adaptive responses of beta cells to sustained glucose stimulation through transcriptional regulation. [Diabetologia (1999) 42: 195±203]

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Glucose induces early growth response gene (Egr-1) expression in pancreatic beta cells

et al. 1999

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“…Preparation of cells took place as previously described [30] with minor modifications. In summary, primary human thyroid cells were obtained as paraadenomatous tissue (assumed to be normal) from thyroidectomies performed at the department of Ear Nose Throat (ENT)-Head and Neck surgery, Rigshospitalet, University of Copenhagen.…”

Section: Methodsmentioning

confidence: 99%

The flame retardant DE-71 (a mixture of polybrominated diphenyl ethers) inhibits human differentiated thyroid cell function in vitro

et al. 2017

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BackgroundNormal thyroid function is essential for general growth and metabolism, but can be affected by endocrine disrupting chemicals (EDCs). Polybrominated diphenyl ethers (PBDEs) have been used worldwide to reduce flammability in different materials and are suspected to be EDCs. The production of the commercial Penta- and OctaBDE mixtures is banned, but DecaBDEs and existing products may leak PBDEs into the environment. Our aim was to investigate the effect of the PentaBDE mixture DE-71 on human thyroid cells in vitro.Materials and methodsPrimary human thyroid cells were obtained as paraadenomatous tissue and cultured in monolayers. The influence of DE-71 on cyclic adenosine monophosphate (cAMP) and thyroglobulin (Tg) production was examined in the culture medium by competitive radioimmunoassay and enzyme-linked immunosorbent assay, respectively. Real-time quantitative PCR analysis of thyroid-specific genes was performed on the exposed cell cultures. PBDE concentrations were determined in cellular and supernatant fractions of the cultures.ResultsDE-71 inhibited Tg-release from TSH-stimulated thyrocytes. At 50 mg/L DE-71, mean Tg production was reduced by 71.9% (range: 8.5–98.7%), and cAMP by 95.1% (range: 91.5–98.8%) compared to controls). Expression of mRNA encoding Tg, TPO and TSHr were significantly inhibited (p<0.0001, p = 0.0079, and p = 0.0002, respectively). The majority of DE-71 added was found in the cell fraction. No cytotoxicity was found.ConclusionsDE-71 inhibited differentiated thyroid cell functions in a two phase response manner and a concentration-dependent inhibition of Tg and cAMP production, respectively, as well as expression of mRNA encoding Tg, TPO and TSHr. Our findings suggest an inhibiting effect of PBDEs on thyroid cells.

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“…To overcome the limitations of inhomogeneous cell cultures, secondary cell cultures could be a solution, but this includes the risk of dedifferentiation and loss of specific properties of the cells as demonstrated in cultures of e.g. thyroid cells [54]. In line with these findings the somatotroph The influence of IL-1β on IL-6/IL-8 release from human GH producing cells in culture expressed as the ratio of IL-6/IL-8 release after IL-1β stimulation compared to the IL-6/IL-8 release in the control groups.…”

Section: Discussionmentioning

confidence: 99%

Interleukin-8 production from human somatotroph adenoma cells is stimulated by interleukin-1β and inhibited by growth hormone releasing hormone and somatostatin

Vindeløv

Hartoft‐Nielsen

Rasmussen

et al. 2011

Growth Hormone & IGF Research

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Human thyroid epithelial cells cultured in monolayers. I. Decreased thyroglobulin and cAMP response to TSH in 12-week-old secondary and tertiary cultures

Cited by 22 publications

References 23 publications

Glucose induces early growth response gene (Egr-1) expression in pancreatic beta cells

Glucose induces early growth response gene (Egr-1) expression in pancreatic beta cells

The flame retardant DE-71 (a mixture of polybrominated diphenyl ethers) inhibits human differentiated thyroid cell function in vitro

Interleukin-8 production from human somatotroph adenoma cells is stimulated by interleukin-1β and inhibited by growth hormone releasing hormone and somatostatin

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