Xanthohumol inhibits cell proliferation and induces apoptosis in human thyroid cells

ABSTRACT An experimental system was established for measuring cell function and proliferation of human thyroid follicles cultured in collagen gel suspended in serum-free medium. Optimal culture conditions were defined and the system was characterized. The human thyrocytes were functional as indicated by their ability to respond to a TSH stimulus (as low as 1–10 μU/ml). in a time- and dose-dependent fashion, with at least a 15-fold increase in iodide uptake and organification, tri-iodothyronine (T3) secretion (demonstrated to derive from de-novo T3 biosynthesis) and cyclic AMP accumulation. Moreover, the same system allowed the measurement of cell proliferation (as indicated by thymidine incorporation and DNA content) following epidermal growth factor (EGF) and phorbol ester challenge under conditions of cell density and medium identical to those for the differentiated functions. The functional responses and cell proliferation were markedly higher compared with those of the same cells in the presence of serum or maintained in monolayer culture. Normal cell polarity, which critically determines functional capacity of thyroid follicles was maintained (as demonstrated by electron microscopy) by the use of collagen gel and serum-free medium. The use of thyroid cells of human origin assumes great importance in view of the wide species differences reported. Cryopreservation of cells rather than the necessity of using freshly derived cells confers greater convenience. The present model system provides a powerful tool for studying human thyroid physiology and pathophysiology. Journal of Endocrinology (1991) 131, 499–506

Download Full-text

Proliferative Activity of Human Thyroid Cells in Various Age Groups and Its Correlation with the Risk of Thyroid Cancer after Radiation Exposure

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jc.2006-0417 ◽

2006 ◽

Vol 91 (7) ◽

pp. 2672-2677 ◽

Cited By ~ 60

Author(s):

Ali G. Saad ◽

Seena Kumar ◽

Elaine Ron ◽

Jay H. Lubin ◽

Jerzy Stanek ◽

...

Keyword(s):

Cell Proliferation ◽

Thyroid Cancer ◽

Proliferative Activity ◽

Age Groups ◽

Inverse Correlation ◽

Immunohistochemical Analysis ◽

Human Thyroid ◽

Ki 67 ◽

Thyroid Cells ◽

Proliferative Rate

Abstract Context: The thyroid gland is vulnerable to the carcinogenic effects of ionizing radiation, and there is a well-documented inverse correlation between thyroid cancer and age at exposure, particularly for ages less than 20 yr. One of the factors responsible for this phenomenon may be more rapid cell proliferation in children. Objective: The objective of this study was to determine the proliferative rate of normal human thyroid cells in different age groups. Design: We used immunohistochemical analysis to determine the Ki-67 proliferative index in 117 thyroid glands obtained at autopsy, including 25 fetal thyroids (11–40 wk gestation), 55 childhood thyroids (0–19 yr), and 37 adult thyroids (20–60 yr). Results: The rate of Ki-67 labeling in the three groups was 7.4 ± 6.10, 0.23 ± 0.15, and 0.08 ± 0.04% respectively, demonstrating an overall trend for diminishing proliferative activity of thyroid cells with increasing age. However, a lack of correlation was noted between the slopes of cancer risk calculated from previous studies of irradiated populations and proliferative rate in the pediatric age intervals of 0–4 and 5–9 yr, suggesting that other factors are likely to be responsible for the particularly high sensitivity to radiation-induced thyroid cancer among the youngest children. Conclusions: Our findings of a general decrease in proliferative activity of thyroid cells with age may explain, at least in part, the higher risks of radiation-related thyroid cancer in children compared with adults. However, the variation in the rate of cell proliferation is unlikely to be responsible entirely for this phenomenon and other factors may also be involved.

Download Full-text

FOXO1 Controls Thyroid Cell Proliferation in Response to TSH and IGF-I and Is Involved in Thyroid Tumorigenesis

Molecular Endocrinology ◽

10.1210/me.2012-1032 ◽

2013 ◽

Vol 27 (1) ◽

pp. 50-62 ◽

Cited By ~ 24

Author(s):

Miguel A. Zaballos ◽

Pilar Santisteban

Keyword(s):

Cell Proliferation ◽

Tumor Cells ◽

Thyroid Tumor ◽

Human Thyroid ◽

Thyroid Cell ◽

Dependent Manner ◽

Thyroid Cells ◽

Differentiated Cells ◽

Igf I ◽

Rat Thyroid

TSH and insulin/IGF-I synergistically induce the proliferation of thyroid cells mainly through the cAMP and phosphatidylinositol 3-kinase (PI3K) pathways. However, the events involved in this cooperative induction remain unknown, and molecules that are potentially controlled by both TSH and IGF-I are interesting candidates as integrators of both stimuli. The finding that the PI3K pathway is frequently activated in thyroid malignancies has attracted attention to this pathway in the thyroid field. One of the targets of PI3K is Forkhead box O (FoxO)-1, a widely expressed transcription factor involved in a variety of cellular processes such as differentiation, proliferation, and apoptosis. Here we show that FoxO1 is highly expressed in differentiated rat thyroid cells and human thyroid tissue compared with human thyroid tumor-derived cells and surgically removed thyroid tumors, in which its expression is reduced. In differentiated cells, TSH/cAMP treatment decreases FoxO1 mRNA and protein levels through proteasome activation, whereas both TSH and IGF-I control FoxO1 localization by promoting a rapid exclusion from the nucleus in an Akt-dependent manner. FoxO1 can control p27KIP1 expression in differentiated and tumor cells of the thyroid. Furthermore, FoxO1 reexpression in tumor cells promotes a decrease in their proliferation rate, whereas FoxO1 interference in differentiated cells increases their proliferation. These data point to an important role of FoxO1 in mediating the effects of TSH and IGF-I on thyroid cell proliferation and provide a link between loss of FoxO1 expression and the uncontrolled proliferation of thyroid tumor cells.

Download Full-text

Modulation of ACE-2 mRNA by inflammatory cytokines in human thyroid cells: a pilot study

Endocrine ◽

10.1007/s12020-021-02807-w ◽

2021 ◽

Author(s):

Francesca Coperchini ◽

Gianluca Ricci ◽

Laura Croce ◽

Marco Denegri ◽

Rubina Ruggiero ◽

...

Keyword(s):

Inflammatory Cytokines ◽

Combination Treatment ◽

Primary Cultures ◽

Human Thyroid ◽

Thyroid Cell ◽

Mrna Levels ◽

Thyroid Cells ◽

Tnf Α ◽

Ifn Γ ◽

Pro Inflammatory Cytokines

Abstract Introduction Angiotensin-converting-enzyme-2 (ACE-2) was demonstrated to be the receptor for cellular entry of SARS-CoV-2. ACE-2 mRNA was identified in several human tissues and recently also in thyroid cells in vitro. Purpose Aim of the present study was to investigate the effect of pro-inflammatory cytokines on the ACE-2 mRNA levels in human thyroid cells in primary cultures. Methods Primary thyroid cell cultures were treated with IFN-γ and TNF-α alone or in combination for 24 h. ACE-2 mRNA levels were measured by RT-PCR. As a control, the levels of IFN-γ inducible chemokine (CXCL10) were measured in the respective cell culture supernatants. Results The mean levels of ACE-2 mRNA increased after treatment with IFN-γ and TNF-α in all the thyroid cell preparations, while the combination treatment did not consistently synergically increase ACE-2-mRNA. At difference, CXCL10 was consistently increased by IFN-γ and synergically further increased by the combination treatment with IFN-γ + TNF-α, with respect to IFN-γ alone. Conclusions The results of the present study show that IFN-γ and, to a lesser extent TNF-α consistently increase ACE-2 mRNA levels in NHT primary cultures. More interestingly, the combined stimulation (proven to be effective according to the synergic effect registered for CXCL10) produces different responses in terms of ACE-2 mRNA modulation. These results would suggest that elevated levels of pro-inflammatory cytokines could facilitate the entering of the virus in cells by further increasing ACE-2 expression and/or account for the different degree of severity of SARS-COV-2 infection. This hypothesis deserves to be confirmed by further specific studies.

Download Full-text

In Vitro Culture of Human Thyroid Cells: Preliminary Findings on the Role of the Pathological Condition of the Donors

Alternatives to Laboratory Animals ◽

10.1177/026119299702500208 ◽

1997 ◽

Vol 25 (2) ◽

pp. 153-160

Author(s):

Francesca Mattioli ◽

Marianna Angiola ◽

Laura Fazzuoli ◽

Francesco Razzetta ◽

Antonietta Martelli

Keyword(s):

Pathological Condition ◽

Primary Cultures ◽

S Phase ◽

Human Thyroid ◽

Thyroid Cells ◽

Toxicological Studies ◽

Predictive Indicator

Although primary cultures of human thyroid cells are used for endocrinological and toxicological studies, until now no attention has been paid toward verifying whether the hormonal conditions to which the gland was exposed in vivo prior to surgery could influence in vitro responses. Our findings suggest that the hormonal situation in vivo cannot be used as a predictive indicator of triiodothyronine and thyroxine release and/or S-phase frequency in vitro, either with or without the addition of bovine thyrotropin.

Download Full-text

Trace Amine-Associated Receptor 1 Trafficking to Cilia of Thyroid Epithelial Cells

Cells ◽

10.3390/cells10061518 ◽

2021 ◽

Vol 10 (6) ◽

pp. 1518

Author(s):

Maria Qatato ◽

Vaishnavi Venugopalan ◽

Alaa Al-Hashimi ◽

Maren Rehders ◽

Aaron D. Valentine ◽

...

Keyword(s):

Cell Surface ◽

Cell Lines ◽

Human Thyroid ◽

Apical Plasma Membrane ◽

Thyroid Cell ◽

Thyroid Cells ◽

Trace Amine ◽

Rat Thyroid

Trace amine-associated receptor 1 (rodent Taar1/human TAAR1) is a G protein-coupled receptor that is mainly recognized for its functions in neuromodulation. Previous in vitro studies suggested that Taar1 may signal from intracellular compartments. However, we have shown Taar1 to localize apically and on ciliary extensions in rodent thyrocytes, suggesting that at least in the thyroid, Taar1 may signal from the cilia at the apical plasma membrane domain of thyrocytes in situ, where it is exposed to the content of the follicle lumen containing putative Taar1 ligands. This study was designed to explore mouse Taar1 (mTaar1) trafficking, heterologously expressed in human and rat thyroid cell lines in order to establish an in vitro system in which Taar1 signaling from the cell surface can be studied in future. The results showed that chimeric mTaar1-EGFP traffics to the apical cell surface and localizes particularly to spherical structures of polarized thyroid cells, procilia, and primary cilia upon serum-starvation. Moreover, mTaar1-EGFP appears to form high molecular mass forms, possibly homodimers and tetramers, in stably expressing human thyroid cell lines. However, only monomeric mTaar1-EGFP was cell surface biotinylated in polarized human thyrocytes. In polarized rat thyrocytes, mTaar1-EGFP is retained in the endoplasmic reticulum, while cilia were reached by mTaar1-EGFP transiently co-expressed in combination with an HA-tagged construct of the related mTaar5. We conclude that Taar1 trafficking to cilia depends on their integrity. The results further suggest that an in vitro cell model was established that recapitulates Taar1 trafficking in thyrocytes in situ, in principle, and will enable studying Taar1 signaling in future, thus extending our general understanding of its potential significance for thyroid autoregulation.

Download Full-text

AN INVESTIGATION OF THE PATHOGENESIS OF HASHIMOTO'S DISEASE BY THYROID TISSUE CULTURE

Journal of Endocrinology ◽

10.1677/joe.0.0200083 ◽

1960 ◽

Vol 20 (2) ◽

pp. 83-NP ◽

Cited By ~ 14

Author(s):

W. J. IRVINE

Keyword(s):

Tissue Culture ◽

Alternative Hypothesis ◽

Thyroid Tissue ◽

Human Thyroid ◽

Rabbit Serum ◽

Thyroid Cell ◽

Thyroid Extract ◽

Thyroid Cells ◽

Hashimoto’S Disease ◽

Hashimoto's Disease

SUMMARY Human thyroid cells were grown in tissue culture in media containing normal human serum, Hashimoto serum, and rabbit sera containing antibodies to purified human thyroglobulin and to crude thyroid extract, respectively. The thyroid cells grew equally well in all media, with the exception of the rabbit serum containing antibodies to crude thyroid extract. Intact thyroid cells obtained from tissue culture failed to fix Hashimoto antibodies in the presence of complement, whereas the constituents of disrupted thyroid cells gave a strongly positive complement-fixation test with Hashimoto serum. It is therefore suggested that the intact thyroid cell is impermeable to complement-fixing Hashimoto antibody. The evidence afforded by the present work adds further weight to the belief that Hashimoto's disease may not be due to a simple auto-immunizing process consequent upon the interaction of thyroid antigen and the known circulating auto-antibodies. Evidence in support of an alternative hypothesis involving 'cell-bound' antibodies with disruption of the follicular basement membrane is discussed.

Download Full-text

The revised 8307 base pair coding sequence of human thyroglobulin transiently expressed in eukaryotic cells

Acta Endocrinologica ◽

10.1530/eje.0.1360508 ◽

1997 ◽

Vol 136 (5) ◽

pp. 508-515 ◽

Cited By ~ 21

Author(s):

Simone A R van de Graaf ◽

Erwin Pauws ◽

Jan J M de Vijlder ◽

Carrie Ris-Stalpers

Keyword(s):

Nucleotide Sequence ◽

Expression System ◽

Thyroid Tissue ◽

Initiation Site ◽

Human Thyroid ◽

Translation Initiation Site ◽

Thyroid Cells ◽

Coding Sequence ◽

Reverse Transcription Pcr ◽

Human Sequence

Abstract We developed a transient transfection system for human thyroglobulin (TG) cDNA in both human thyroid cells and in COS-1 cells. Four overlapping TG cDNA fragments were amplified by reverse transcription-PCR from RNA of normal thyroid tissue. The most 5′ fragment includes the natural translation initiation site and the sequence encoding the signal peptide (SP). After subcloning, the nucleotide sequence was determined and compared with the published human sequence, resulting in the detection of 30 nucleotide variations. For validation purposes, all variations were screened in 6–12 normal human alleles. Twenty-one were present in all screened alleles and have to be revised in the published nucleotide sequence. Since one variation concerns a triplet insertion, the coding sequence of the mature human thyroglobulin is 8307 nucleotides encoding 2750 amino acids. The TG cDNA constructs were transiently transfected in HTori 3 and COS-1 cells and protein expression was detected using a polyclonal anti-human-TG on fixed cells and after SDS-PAGE. In both cell-lines all four TG protein fragments were expressed. The mannose structures detected on the proteins by lectins and localization after expression in the cells suggest that only the N-terminal TG fragment (containing the SP) is directed to the endoplasmatic reticulum but is unable to reach the Golgi complex. The described expression system in human thyrocytes will be a helpful tool in studying the structure–function relationship of human TG in thyroid hormonogenesis. European Journal of Endocrinology 136 508–515

Download Full-text