scholarly journals Progesterone Upregulates Gene Expression in Normal Human Thyroid Follicular Cells

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Ana Paula Santin Bertoni ◽  
Ilma Simoni Brum ◽  
Ana Caroline Hillebrand ◽  
Tania Weber Furlanetto
Keyword(s):  
Mrna Expression ◽  
Cell Function ◽  
Human Thyroid ◽  
Thyroid Cell ◽  
Sodium Iodide Symporter ◽  
Follicular Cells ◽  
Ki 67 ◽  
Thyroid Cells ◽  
Female Sex Hormones ◽  
Thyroid Follicular Cells

Thyroid cancer and thyroid nodules are more prevalent in women than men, so female sex hormones may have an etiological role in these conditions. There are no data about direct effects of progesterone on thyroid cells, so the aim of the present study was to evaluate progesterone effects in the sodium-iodide symporterNIS, thyroglobulinTG, thyroperoxidaseTPO, andKI-67genes expression, in normal thyroid follicular cells, derived from human tissue.NIS,TG,TPO, andKI-67mRNA expression increased significantly after TSH 20 μUI/mL, respectively: 2.08 times,P<0.0001; 2.39 times,P=0.01; 1.58 times,P=0.0003; and 1.87 times,P<0.0001. In thyroid cells treated with 20 μUI/mL TSH plus 10 nM progesterone, RNA expression ofNIS,TG, andKI-67genes increased, respectively: 1.78 times,P<0.0001; 1.75 times,P=0.037; and 1.95 times,P<0.0001, andTPOmRNA expression also increased, though not significantly (1.77 times,P=0.069). These effects were abolished by mifepristone, an antagonist of progesterone receptor, suggesting that genes involved in thyroid cell function and proliferation are upregulated by progesterone. This work provides evidence that progesterone has a direct effect on thyroid cells, upregulating genes involved in thyroid function and growth.

scholarly journals Detection of SARS-COV-2 receptor ACE-2 mRNA in thyroid cells: a clue for COVID-19-related subacute thyroiditis

2020 ◽  
Author(s):  
M. Rotondi ◽  
F. Coperchini ◽  
G. Ricci ◽  
M. Denegri ◽  
L. Croce ◽  
...  
Keyword(s):  
Primary Cultures ◽  
Thyroid Tissue ◽  
Subacute Thyroiditis ◽  
Human Thyroid ◽  
Thyroid Cell ◽  
Type I ◽  
Rt Pcr ◽  
Follicular Cells ◽  
Thyroid Cells ◽  
Tissue Samples

Abstract Purpose SARS-COV-2 is a pathogenic agent belonging to the coronavirus family, responsible for the current global world pandemic. Angiotensin-converting enzyme 2 (ACE-2) is the receptor for cellular entry of SARS-CoV-2. ACE-2 is a type I transmembrane metallo-carboxypeptidase involved in the Renin-Angiotensin pathway. By analyzing two independent databases, ACE-2 was identified in several human tissues including the thyroid. Although some cases of COVID-19-related subacute thyroiditis were recently described, direct proof for the expression of the ACE-2 mRNA in thyroid cells is still lacking. Aim of the present study was to investigate by RT-PCR whether the mRNA encoding for ACE-2 is present in human thyroid cells. Methods RT-PCR was performed on in vitro ex vivo study on thyroid tissue samples (15 patients undergoing thyroidectomy for benign thyroid nodules) and primary thyroid cell cultures. Results The ACE-2 mRNA was detected in all surgical thyroid tissue samples (n = 15). Compared with two reporter genes (GAPDH: 0.052 ± 0.0026 Cycles−1; β-actin: 0.044 ± 0.0025 Cycles−1; ACE-2: 0.035 ± 0.0024 Cycles−1), the mean level of transcript expression for ACE-2 mRNA was abundant. The expression of ACE-2 mRNA in follicular cells was confirmed by analyzing primary cultures of thyroid cells, which expressed the ACE-2 mRNA at levels similar to tissues. Conclusions The results of the present study demonstrate that the mRNA encoding for the ACE-2 receptor is expressed in thyroid follicular cells, making them a potential target for SARS-COV-2 entry. Future clinical studies in patients with COVID-19 will be required for increase our understanding of the thyroid repercussions of SARS-CoV-2 infection.

Expression of an intercellular adhesion molecule, ICAM-1, by human thyroid cells

1989 ◽  
Vol 122 (1) ◽  
pp. 185-NP ◽  
Author(s):  
A. P. Weetman ◽  
S. Cohen ◽  
M. W. Makgoba ◽  
L. K. Borysiewicz
Keyword(s):  
Adhesion Molecule ◽  
Intercellular Adhesion ◽  
Intercellular Adhesion Molecule ◽  
Human Thyroid ◽  
Accessory Cell ◽  
Graves Disease ◽  
Follicular Cells ◽  
Thyroid Cells ◽  
Hla Dr ◽  
Thyroid Follicular Cells

ABSTRACT Intercellular adhesion molecule-1 (ICAM-1), hitherto identified on activated B cells, macrophages, dendritic cells, endothelia and certain epithelial cells, serves as a ligand for the lymphocyte function-associated antigen-1 (LFA-1). ICAM-1 binding by LFA-1 enhances the efficiency of lymphocyte-target cell and lymphocyte-accessory cell interactions. We have investigated the in-vitro expression of ICAM-1 by cultured thyroid cells from five patients with Graves' disease using indirect immunofluorescence analysis, and found that 30 ± 11% (mean ± s.d.) of cells were ICAM-1 positive under basal conditions. The proportion of cells which were ICAM-1 positive and the amount of ICAM-1 per cell (assessed by fluorescence intensity) were both increased in all cases by the cytokines γ-interferon, interleukin-1 and tumour necrosis factor. Immunohistochemical analysis of frozen sections from thyroidectomy specimens demonstrated ICAM-1 on thyroid follicular cells in areas of lymphocytic infiltration in patients with Graves' disease (n = 2) or Hashimoto's thyroiditis (n = 2). ICAM-1 was not found in specimens from a patient with a toxic multinodular goitre or a patient with Graves' disease without focal lymphocytic accumulation. These results suggest that the thyroid epithelium may express ICAM-1 as well as major histocompatibility complex class II antigens, such as HLA-DR, in response to locally synthesized cytokines. The enhanced expression of ICAM-1 may render these cells more susceptible as targets for lymphocytemediated cytotoxicity, and together with HLA-DR antigen expression may increase the accessory cell capability of the thyroid follicular cells. Journal of Endocrinology (1989) 122, 185–191

Relationship between growth and function of human thyroid cells in culture

1986 ◽  
Vol 108 (3) ◽  
pp. 393-398 ◽  
Author(s):  
C. A. Ollis ◽  
R. Davies ◽  
D. S. Munro ◽  
S. Tomlinson
Keyword(s):  
Cell Growth ◽  
Growth Response ◽  
Cell Function ◽  
Human Thyroid ◽  
Thyroid Cell ◽  
Thyroid Cells ◽  
Cells In Culture ◽  
Epidermal Growth ◽  
And Function ◽  
Human Thyroid Cell

ABSTRACT Subconfluent human thyroid cells in monolayer, isolated from thyrotoxic tissue or non-toxic goitres obtained at surgery, responded to the addition of epidermal growth factor (EGF) with an increase in cell growth as measured by increased incorporation of [3H]thymidine into trichloroacetic acid-precipitable material. The growth response to EGF was concentration-dependent and the characteristics of the responses were the same using EGF from murine or human sources. With concentrations which stimulated growth, EGF was found to inhibit human thyroid cell function as measured by the release of radioimmunoassayable tri-iodothyronine into the incubation medium. Thyrotrophin (TSH) was also found to stimulate human thyroid cell growth but at concentrations far lower than those used to stimulate thyroid cell function in this system. The effect of EGF on the differentiating action of TSH on human thyroid cells in culture was also investigated; the association of thyroid cells into two-dimensional follicular structures produced by the incubation of thyroid cells at a high cell density with TSH was found to be inhibited by the addition of EGF. J. Endocr. (1986) 108, 393–398

Validation of Immunohistochemistry for Canine Proteins Involved in Thyroid Iodine Uptake and Their Expression in Canine Follicular Cell Thyroid Carcinomas (FTCs) and FTC-Derived Organoids

2021 ◽  
pp. 030098582110188
Author(s):  
Jana Jankovic ◽  
Martina Dettwiler ◽  
Martin González Fernández ◽  
Eve Tièche ◽  
Kerstin Hahn ◽  
...  
Keyword(s):  
Thyroid Gland ◽  
Western Blot ◽  
Follicular Cell ◽  
Iodine Uptake ◽  
Human Thyroid ◽  
Thyroid Peroxidase ◽  
Thyroid Cell ◽  
Follicular Cells ◽  
Thyroid Cells ◽  
Primary Tumors

Thyrotropin receptor (TSHR), sodium iodide symporter (NIS), pendrin, and thyroid peroxidase (TPO) are essential for the uptake of iodine by follicular thyroid cells. The aim of this study was to establish immunohistochemistry (IHC) protocols for TSHR, NIS, pendrin, and TPO in canine tissues and characterize their expression in organoids derived from canine follicular cell thyroid carcinoma (FTC) and in the respective primary tumors. This constitutes a fundamental step to establish organoids as a model to study the uptake of iodine in canine FTC. Commercially available antibodies directed against human proteins were selected. Antibody specificity was confirmed by western blot using lysates of the HTori-3 human thyroid cell line and healthy canine thyroid gland. IHC was validated using HTori-3 cells and a set of canine normal tissues including healthy thyroid gland. The expression of TSHR, NIS, pendrin, and TPO was evaluated in 3 organoid lines derived from FTC and respective primary tumors. All 4 antibodies produced specific bands by western blot and cytoplasmic labeling in follicular cells by IHC in both human HTori-3 cells and canine thyroid gland. NIS also showed basolateral membrane immunolabeling in follicular cells. All 4 proteins were highly expressed in organoids derived from FTC. The expression was similar or higher compared to the primary tumors. The results of this study characterize organoids derived from canine FTC as a suitable in vitro model to investigate iodine uptake, opening new research possibilities in the field of canine thyroid cancer therapy.

FK506 inhibits phytohemagglutinin-, but not interferon-γ-, induced HLA-DR antigen expression and accessory cell function on primary cultured human thyroid cells

1993 ◽  
Vol 129 (6) ◽  
pp. 579-584 ◽  
Author(s):  
Masayuki Sato ◽  
Yuji Hiromatsu ◽  
Kiyoko Tanaka ◽  
Noriko Ishisaka ◽  
Kyohei Nonaka
Keyword(s):  
Cell Function ◽  
Antigen Expression ◽  
Interferon Γ ◽  
Human Thyroid ◽  
Accessory Cell ◽  
Thyroid Cell ◽  
Dependent Manner ◽  
Thyroid Cells ◽  
Hla Dr ◽  
Ifn Γ

We investigated the effects of FK506, a novel immunosuppressive agent, on the phytohemagglutinin (PHA) or interferon-γ (IFN-γ)-induced expression of HLA-DR antigen, accessory cell function and proliferation of primary cultured human thyroid cells. Primary cultured thyroid cells from patients with Graves' disease were incubated for 3 days with PHA in concentrations in the range 1–50 mg/l or with 200 kU/l of IFN-γ, in the presence or absence of FK506. The surface expression of HLA-DR antigen was measured by flow cytometry. Accessory cell function of thyroid cells was assessed by the incorporation of [3H]thymidine to T cells in the presence of 0.1–1.0 μg/l staphylococcus enterotoxin B (SEB). The proliferation of thyroid cells was determined from [3H]thymidine incorporation assays. FK506 inhibited the induction of HLA-DR antigen expression by PHA on thyroid cells in a dose-dependent manner, but did not inhibit that by IFN-γ. Polyclonal anti-IFN-γ antibody partly inhibited the PHA-induced HLA-DR antigen expression on thyroid cells. Phytohemagglutinin enhanced the SEB-mediated accessory cell function of thyroid cells. FK506 inhibited the accessory cell function induced by PHA. FK506 alone did not directly affect the thyroid cell proliferation, although it ameliorated the thyroid cell growth suppressed by PHA. Our data suggest that FK506 suppresses the HLA-DR antigen expression induced by PHA and the subsequent accessory cell function on thyroid cells via the inhibition of T lymphocytes present in the primary culture.

scholarly journals Sphingolipid-Cholesterol Domains (Lipid Rafts) in Normal Human and Dog Thyroid Follicular Cells Are Not Involved in Thyrotropin Receptor Signaling

Endocrinology ◽  
2004 ◽  
Vol 145 (3) ◽  
pp. 1464-1472 ◽  
Author(s):  
M. J. Costa ◽  
Y. Song ◽  
P. Macours ◽  
C. Massart ◽  
M. C. Many ◽  
...  
Keyword(s):  
Primary Cultures ◽  
Tsh Receptor ◽  
Human Thyroid ◽  
Membrane Domains ◽  
Follicular Cells ◽  
Thyroid Cells ◽  
Caveolin 1 ◽  
Thyroid Follicular Cells ◽  
First Time ◽  
Camp Cascade

Abstract Partition of signaling molecules in sphingolipid-cholesterol-enriched membrane domains, among which are the caveolae, may contribute to signal transduction efficiency. In normal thyroid, nothing is known about a putative TSH/cAMP cascade compartmentation in caveolae or other sphingolipid-cholesterol-enriched membrane domains. In this study we show for the first time that caveolae are present in the apical membrane of dog and human thyrocytes: caveolin-1 mRNA presence is demonstrated by Northern blotting in primary cultures and that of the caveolin-1 protein by immunohistochemistry performed on human thyroid tissue. The TSH receptor located in the basal membrane can therefore not be located in caveolae. We demonstrate for the first time by biochemical methods the existence of sphingolipid-cholesterol-enriched domains in human and dog thyroid follicular cells that contain caveolin, flotillin-2, and the insulin receptor. We assessed a possible sphingolipid-cholesterol-enriched domains compartmentation of the TSH receptor and the α- subunit of the heterotrimeric Gs and Gq proteins using two approaches: Western blotting on detergent-resistant membranes isolated from thyrocytes in primary cultures and the influence of 10 mm methyl-β-cyclodextrin, a cholesterol chelator, on basal and stimulated cAMP accumulation in intact thyrocytes. The results from both types of experiments strongly suggest that the TSH/cAMP cascade in thyroid cells is not associated with sphingolipid-cholesterol-enriched membrane domains.

Inhibitory effect of dihydrotestosterone on human thyroid cell growth

1996 ◽  
Vol 151 (2) ◽  
pp. 185-194 ◽  
Author(s):  
R Rossi ◽  
M C Zatelli ◽  
P Franceschetti ◽  
I Maestri ◽  
E Magri ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Growth ◽  
Primary Cultures ◽  
Inhibitory Effect ◽  
Human Thyroid ◽  
Growth Fraction ◽  
Thyroid Cell ◽  
Mrna Levels ◽  
Follicular Cells ◽  
Thyroid Follicular Cells

Abstract Sex steroid-binding activities have been identified by several authors in normal and pathological thyroids and the expression of the canonic androgen receptor (AR) has recently been demonstrated in human thyroid follicular cells. In order to assess what influence, if any, androgen exposure has on thyroid cell growth, the effect of dihydrotestosterone (DHT) on [3H]thymidine (thy) incorporation and cell proliferation was investigated in thyroid follicular cells in vitro. In a primary culture of goitrous cells, DHT induced a significant reduction of [3H]thy incorporation at concentrations ranging from 10−12 to 10−8 m, with a more pronounced effect at 10−9 m. At this concentration, the inhibitory effect was evident after both 24 and 48 h of treatment and in various types of primary thyroid cell cultures. In goitrous cells, the DHT-induced decrease of [3H]thy was associated with a reduction of expression of the proliferation-associated nuclear Ki-67 antigen, a protein commonly used to assess cell growth fraction. In TPC cells, an AR-positive thyroid papillary carcinoma cell line, DHT at concentrations between 10−12 and 10−8 m significantly decreased the growth rate. DHT (10−9 m) produced an approximately 50–60% inhibition of cell proliferation and the antiandrogen cyproterone acetate was capable of reversing such effects. The DHT-induced reduction of TPC cell proliferation was associated with a significant reduction of c-myc RNA levels. Thyroperoxidase mRNA levels and thyroglobulin production were not reduced by androgen in primary cultures of goitrous cells. In conclusion, our results indicated that androgens may have a role in this gland by reducing the proliferation, but not the function, of follicular cells. Journal of Endocrinology (1996) 151, 185–194

scholarly journals An Amphotropic Retroviral Vector Expressing a Mutant gsp Oncogene: Effects on Human Thyroid Cells in Vitro1

1997 ◽  
Vol 82 (8) ◽  
pp. 2702-2709 ◽  
Author(s):  
M. Ivan ◽  
M. Ludgate ◽  
V. Gire ◽  
J. A. Bond ◽  
D. Wynford-Thomas
Keyword(s):  
Retroviral Vector ◽  
Human Thyroid ◽  
Nodule Formation ◽  
Target Cells ◽  
Follicular Cells ◽  
Morphological Response ◽  
Thyroid Cells ◽  
Significant Difference ◽  
Thyroid Follicular Cells ◽  
Normal Human

Point mutations of the gsp protooncogene (encoding theα -subunit of the Gs protein) that constitutively activate the cAMP signaling pathway are a common feature of and a plausible causative mechanism for thyroid hyperfunctioning adenomas (hot nodules). To investigate the extent to which mutant gsp acting alone can induce proliferation of thyroid follicular cells, we generated an amphotropic retroviral vector (based on the pBABE-neo plasmid and psi-CRIP packaging line) to permit stable introduction of a hemagglutinin-tagged Gln227→Leu mutant gsp gene into normal human thyrocytes in vitro. The biological activity of the vector was confirmed by detection of HA-tagged Gsp protein expression and induction of cAMP synthesis in selected target cells. Normal human thyroid follicular cells in primary monolayer culture were infected with the gsp retroviral vector or with corresponding vectors expressing mutant H-ras or neo only as positive and negative controls, respectively. Although, as before, mutant ras generated 10–20 well differentiated epithelial colonies/dish of 105 infected cells, with an average lifespan of 15–20 population doublings, only small groups of no more than 15–50 differentiated thyrocytes were observed with the gsp vector. In addition to standard conditions (10% FCS), infections were performed in reduced serum (1% FCS, TSH, and insulin), in the presence of isobutylylmethylxanthine, or in the presence of agents capable of closing gap junctions, with no significant difference in outcome. Although little or no proliferative response was observed regardless of the conditions, there was clear evidence of morphological response (rearrangement of the actin cytoskeleton and increased cell size). The results suggest that gsp mutation may not be a sufficient proliferogenic stimulus by itself to account for hot nodule formation.

scholarly journals MAPK Inhibition Requires Active RAC1 Signaling to Effectively Improve Iodide Uptake by Thyroid Follicular Cells

Cancers ◽  
2021 ◽  
Vol 13 (22) ◽  
pp. 5861
Author(s):  
Márcia Faria ◽  
Rita Domingues ◽  
Maria João Bugalho ◽  
Paulo Matos ◽  
Ana Luísa Silva
Keyword(s):  
Positive Impact ◽  
Mapk Pathway ◽  
Functional Expression ◽  
Braf V600e ◽  
Thyroid Cell ◽  
Follicular Cells ◽  
Iodide Uptake ◽  
Thyroid Cells ◽  
Mek Inhibitors ◽  
Thyroid Follicular Cells

The Sodium/Iodide Symporter (NIS) is responsible for the active transport of iodide into thyroid follicular cells. Differentiated thyroid carcinomas (DTCs) usually preserve the functional expression of NIS, allowing the use of radioactive iodine (RAI) as the treatment of choice for metastatic disease. However, a significant proportion of patients with advanced forms of TC become refractory to RAI therapy and no effective therapeutic alternatives are available. Impaired iodide uptake is mainly caused by the defective functional expression of NIS, and this has been associated with several pathways linked to malignant transformation. MAPK signaling has emerged as one of the main pathways implicated in thyroid tumorigenesis, and its overactivation has been associated with the downregulation of NIS expression. Thus, several strategies have been developed to target the MAPK pathway attempting to increase iodide uptake in refractory DTC. However, MAPK inhibitors have had only partial success in restoring NIS expression and, in most cases, it remained insufficient to allow effective treatment with RAI. In a previous work, we have shown that the activity of the small GTPase RAC1 has a positive impact on TSH-induced NIS expression and iodide uptake in thyroid cells. RAC1 is a downstream effector of NRAS, but not of BRAF. Therefore, we hypothesized that the positive regulation induced by RAC1 on NIS could be a relevant signaling cue in the mechanism underlying the differential response to MEK inhibitors, observed between NRAS- and BRAF-mutant tumors. In the present study, we found that the recovery of NIS expression induced through MAPK pathway inhibition can be enhanced by potentiating RAC1 activity in thyroid cell systems. The negative impact on NIS expression induced by the MAPK-activating alterations, NRAS Q61R and BRAF V600E, was partially reversed by the presence of the MEK 1/2 inhibitors AZD6244 and CH5126766. Notably, the inhibition of RAC1 signaling partially blocked the positive impact of MEK inhibition on NIS expression in NRAS Q61R cells. Conversely, the presence of active RAC1 considerably improved the rescue of NIS expression in BRAF V600E thyroid cells treated with MEK inhibitors. Overall, our data support an important role for RAC1 signaling in enhancing MAPK inhibition in the context of RAI therapy in DTC, opening new opportunities for therapeutic intervention.

Studies of catecholamine effect on cyclic AMP in human cultured thyroid cells: their interaction with thyrotrophin receptor

1983 ◽  
Vol 102 (1) ◽  
pp. 62-67 ◽  
Author(s):  
Roberto S. Toccafondi ◽  
Maria Luisa Brandi ◽  
Carlo M. Rotella ◽  
Roberto Zonefrati
Keyword(s):  
Cyclic Amp ◽  
Adrenergic Receptors ◽  
Human Thyroid ◽  
Adrenergic Nerve ◽  
Camp Accumulation ◽  
Follicular Cells ◽  
Functional Responses ◽  
Thyroid Cells ◽  
Thyroid Follicular Cells ◽  
Bovine Tsh

Abstract. Even though adrenergic nerve terminals between and around thyroid follicles and catecholamine stimulation of thyroid adenylate cyclase have been reported, there is no uniform concept on catecholamine interaction with thyrotrophin (TSH) receptors. Therefore, the effect of catecholamines on TSH-stimulated cyclic AMP (cAMP) accumulation in human follicular thyroid cells has been investigated, to thus eliminating the extrathyroidal actions of catecholamines. Epinephrine, norepinephrine and isoproterenol appeared to be rapid and potent stimulators of intracellular cAMP accumulation, the half maximum increase doses being 4 × 10−7m, 1 × 10−5m and 5 × 10−7m, respectively. While propranolol (1 × 10−5m) prevented the stimulatory effect of catecholamines and failed to inhibit the effect of bovine TSH, phentolamine (1 × 10−5m) enhanced the potency of norepinephrine and bovine TSH, leaving that of epinephrine unchanged. The effects of epinephrine (2 × 10−8m) and isoproterenol (2 × 10−8m) were additive to that of bovine TSH (0.5 mU/ml), but the effect of simultaneous stimulation with norepinephrine (5 × 10−7m) and bovine TSH (0.5 mU/ml) was lower than expected. Prenalterol, a selective β1-agonist, did not stimulate cAMP accumulation, while terbutaline, a selective β2-agonist, exerted a potent stimulation. Metoprolol, a selective β1-adrenergic blocker, did not affect the response of thyroid follicular cells to isoproterenol. These results demonstrate the existence of β-adrenergic receptors in human thyroid follicular cells, mainly of the type β2, apparently not correlated with TSH receptor. The existence of α-adrenergic receptors which counter-regulate TSH functional responses in human thyroid follicular cells is suggested.

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