Epidermal growth factor (GGF) and tumor promoter 12-0-tetradecanoylphorbol 13-acetate (TPA) stimulate PG synthesis and thymidine incorporation in cultured porcine thyroid cells

1987 ◽  
Vol 143 (3) ◽  
pp. 880-884 ◽  
Author(s):  
Nobuyuki Takasu ◽  
Seiya Sato ◽  
Takashi Yamada ◽  
Yoshifusa Shimizu
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Thymidine Incorporation ◽  
Tumor Promoter ◽  
Thyroid Cells ◽  
Epidermal Growth

scholarly journals Tri-iodothyronine induces proliferation in cultured bovine thyroid cells: evidence for the involvement of epidermal growth factor-associated tyrosine kinase activity

2000 ◽  
Vol 166 (1) ◽  
pp. 173-182 ◽  
Author(s):  
M Di Fulvio ◽  
AH Coleoni ◽  
CG Pellizas ◽  
AM Masini-Repiso
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Western Blot ◽  
Blot Analysis ◽  
Thymidine Incorporation ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Thyroid Cells ◽  
Epidermal Growth ◽  
Bovine Thyroid

The effects of the tri-iodothyronine (T(3)) secreted by thyroid cells on the growth of the thyrocyte are poorly known. In this study we analyzed the effects of T(3) on the proliferation of bovine thyroid follicles in primary culture previously depleted of endogenous T(3). Cellular deoxiribonucleic acid (DNA) synthesis, determined by [(3)H]thymidine incorporation, was stimulated by T(3) (0.1-5.0 nM) for 24 h in a concentration-dependent fashion with a maximal effect at 1.0 nM T(3) (P<0.01). This T(3) action was time-dependent when assayed from 12 to 72 h. The induction of mitogenic activity was corroborated by the increase in proliferating cell nuclear antigen (PCNA) measured by Western blot analysis. PCNA increased after treatment with T(3) (0.1-5.0 nM) in a concentration-dependent manner. Since T(3) modifies the activity of growth factors whose actions are mainly mediated by tyrosine kinase (TK) activation in diverse cellular types, we assayed the effects of genistein, a general TK inhibitor, and tyrphostin A25, a specific epidermal growth factor (EGF)-receptor (EGFR)-dependent TK activity inhibitor, on the proliferative effects of T(3). The T(3)-induced [(3)H]thymidine incorporation was inhibited by both agents in a concentration-dependent manner. A significant increase in the total TK activity measured in cellular protein extracts was induced by 0.5 and 1.0 nM T(3) (P<0.001). Tyrosine phosphorylation of the EGFR was also stimulated by T(3) (P<0.001) with no change in the EGFR expression as determined by Western blot analysis. Both, the T(3)-stimulated [(3)H]thymidine incorporation and the TK activity were inhibited by a anti-mouse EGF antibody. These results lead us to propose that T(3) could operate as a proliferative agent in bovine thyroid cells through a mechanism involving an autocrine/paracrine EGF/EGFR-dependent regulation.

Tumour promoter 12-O-tetradecanoylphorbol 13-acetate and epidermal growth factor stimulate proliferation and inhibit differentiation of porcine thyroid cells in primary culture

1987 ◽  
Vol 113 (3) ◽  
pp. 485-487 ◽  
Author(s):  
N. Takasu ◽  
Y. Shimizu ◽  
T. Yamada
Keyword(s):  
Growth Factor ◽  
Thyroid Hormone ◽  
Epidermal Growth Factor ◽  
Primary Culture ◽  
Thymidine Incorporation ◽  
Biological Activities ◽  
Early Step ◽  
Thyroid Cells ◽  
Hormone Synthesis ◽  
Epidermal Growth

ABSTRACT 12-O-Tetradecanoylphorbol 13-acetate (TPA) is a potent tumour promoter and shows several biological activities of epidermal growth factor (EGF). EGF and TPA stimulated proliferation and inhibited differentiation of porcine thyroid cells in primary culture. They also stimulated [3H]thymidine incorporation and inhibited an early step in thyroid hormone synthesis (iodine organification). The results indicate that EGF and TPA switch the developmental course of porcine thyroid cells from differentiation to proliferation. J. Endocr. (1987) 113, 485–487

An AP1-binding site in the c-fos gene can mediate induction by epidermal growth factor and 12-O-tetradecanoyl phorbol-13-acetate

1989 ◽  
Vol 9 (3) ◽  
pp. 1327-1331
Author(s):  
T M Fisch ◽  
R Prywes ◽  
R G Roeder
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Binding Site ◽  
Tumor Promoter ◽  
Consensus Binding Site ◽  
Sequence Element ◽  
Fos Protein ◽  
Sequence Elements ◽  
Epidermal Growth ◽  
Serum Response

We have demonstrated that two sequence elements in the c-fos promoter can mediate the response of the gene to epidermal growth factor and the tumor promoter 12-O-tetradecanoyl phorbol-13-acetate (TPA). The first is the previously described serum response element. The second is a sequence element highly homologous to the consensus binding site for the HeLa cell transcription factor AP1. Although recent reports have shown that fos protein binds to AP1-binding sites through an interaction with AP1 protein and have raised the speculation that fos protein may negatively regulate expression of the c-fos gene via this interaction, we found no role for the AP1 consensus homology in the downregulation of c-fos expression following induction by epidermal growth factor and TPA.

Effects of epidermal growth factor and 12-O-tetradecanoylphorbol-13-acetate on metabolism of the epidermal growth factor receptor in normal human fibroblasts

1984 ◽  
Vol 4 (9) ◽  
pp. 1718-1724
Author(s):  
S J Decker
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Half Life ◽  
Egf Receptor ◽  
Human Fibroblasts ◽  
Tumor Promoter ◽  
Minor Amount ◽  
Detectable Effect ◽  
Normal Human ◽  
Epidermal Growth

The biosynthesis, phosphorylation, and degradation of the epidermal growth factor (EGF) receptor were examined in normal human fibroblasts. The receptor was initially synthesized as an Mr = 160,000 immature form which matured to an Mr = 170,000 form in a monensin-sensitive manner. Tunicamycin treatment led to the accumulation of an Mr = 130,000 protein. The receptor was phosphorylated on serine and threonine residues in normally growing and quiescent cells, and treatment with EGF or the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) resulted in a two- to threefold increase in receptor-bound phosphate. EGF increased the amount of phosphoserine and phosphothreonine and caused the appearance of a minor amount of phosphotyrosine. TPA increased the levels of phosphoserine and phosphothreonine exclusively. Prior treatment with TPA inhibited the EGF-dependent appearance of phosphotyrosine in the receptor. Analysis of tryptic phosphopeptides revealed that six of the seven major peptides were common to the receptor from cells treated with EGF or TPA. EGF strongly stimulated [3H]thymidine incorporation in confluent cells, increased final saturation density three to fourfold, and increased whole-cell levels of phosphotyrosine about threefold. Treatment of cells with TPA before addition of EGF inhibited all three of these EGF-dependent responses. EGF also decreased the receptor half-life from 15 h to 1 h, but this was not inhibited by TPA. TPA alone had no detectable effect on the receptor half-life.

scholarly journals Effects of epidermal growth factor and 12-O-tetradecanoylphorbol-13-acetate on metabolism of the epidermal growth factor receptor in normal human fibroblasts.

1984 ◽  
Vol 4 (9) ◽  
pp. 1718-1724 ◽  
Author(s):  
S J Decker
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Half Life ◽  
Egf Receptor ◽  
Human Fibroblasts ◽  
Tumor Promoter ◽  
Minor Amount ◽  
Detectable Effect ◽  
Normal Human ◽  
Epidermal Growth

The biosynthesis, phosphorylation, and degradation of the epidermal growth factor (EGF) receptor were examined in normal human fibroblasts. The receptor was initially synthesized as an Mr = 160,000 immature form which matured to an Mr = 170,000 form in a monensin-sensitive manner. Tunicamycin treatment led to the accumulation of an Mr = 130,000 protein. The receptor was phosphorylated on serine and threonine residues in normally growing and quiescent cells, and treatment with EGF or the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) resulted in a two- to threefold increase in receptor-bound phosphate. EGF increased the amount of phosphoserine and phosphothreonine and caused the appearance of a minor amount of phosphotyrosine. TPA increased the levels of phosphoserine and phosphothreonine exclusively. Prior treatment with TPA inhibited the EGF-dependent appearance of phosphotyrosine in the receptor. Analysis of tryptic phosphopeptides revealed that six of the seven major peptides were common to the receptor from cells treated with EGF or TPA. EGF strongly stimulated [3H]thymidine incorporation in confluent cells, increased final saturation density three to fourfold, and increased whole-cell levels of phosphotyrosine about threefold. Treatment of cells with TPA before addition of EGF inhibited all three of these EGF-dependent responses. EGF also decreased the receptor half-life from 15 h to 1 h, but this was not inhibited by TPA. TPA alone had no detectable effect on the receptor half-life.

134. EPIDERMAL GROWTH FACTOR RECEPTOR/MAPK3/1 PATHWAY CROSS-TALK ENABLES GROWTH DIFFERENTIATION FACTOR 9 TO SIGNAL THROUGH SMAD2/3 IN MOUSE GRANULOSA CELLS

2009 ◽  
Vol 21 (9) ◽  
pp. 53
Author(s):  
M. Sasseville ◽  
L. J. Ritter ◽  
T. Nguyen ◽  
D. G. Mottershead ◽  
D. L. Russell ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Granulosa Cells ◽  
Cross Talk ◽  
Egf Receptor ◽  
Thymidine Incorporation ◽  
Differentiation Factor ◽  
Growth Differentiation Factor 9 ◽  
Epidermal Growth ◽  
Receptor Inhibitor

Oocyte-secreted growth differentiation factor 9 (GDF9) plays a critical role throughout folliculogenesis. It has been shown to control many functions of granulosa cells, including gene expression, steroidogenesis and proliferation. This study investigates the cellular requirements that allow GDF9 to act on granulosa cells. Our results showed that GDF9 (20 ng/ml)-stimulated mouse granulosa cells 3H-thymidine incorporation was inhibited by a type 1 receptor Alk4/5/7 inhibitor (SB431542, 5 μM), by an epidermal growth factor (EGF) receptor inhibitor (AG1478, 5μM) and a MEK1 inhibitor (U0126, 10 μM). Interestingly, activin A- and TGFβ-stimulated 3H-thymidine incorporation shared similar inhibitor sensitivity. Moreover, when denuded oocytes were used as the mitogenic agent, SB431542, AG1478 and U0126 all prevented the increase in 3H-thymidine incorporation. Oocyte-stimulated 3H-thymidine incorporation in secondary follicles and cumulus-oocyte complexes were also sensitive to Alk4/5/7, EGF receptor and MEK1 inhibition. Basal and EGF-stimulated levels of phopho-MAPK3/1 were inhibited by using the EGF receptor inhibitor, but were not affected by inhibition of Alk4/5/7 or by adding GDF9 in granulosa cells. Using granulosa cells transfected with a SMAD3-luciferase reporter construct, GDF9-stimulated SMAD3 response could be inhibited by Alk4/5/7, EGFR and MEK1 inhibitors. Genes involved in cumulus cells expansion (Ptx3 and Has2) were upregulated in granulosa cells by co-culturing with denuded oocytes and that upregulation was inhibited by Alk4/5/7 as well as by EGF receptor inhibition. These results suggest that TGFβ superfamily members signalling through Smad2/3 share a common requirement of EGF receptor-dependant phospho-MAPK3/1 throughout folliculogenesis. These results strongly suggest that, apart from its role in the transmission of the ovulatory LH signal within the ovarian follicle, EGF receptor pathway might serve as modulators of GDF9 action on granulosa cells. Hence the interaction between endocrine and oocyte signalling may be mediated at the level of MAPK and Smad2/3 cross-talk in granulosa cells.

The proliferative responses of porcine thyroid follicular cells to epidermal growth factor and thyrotrophin reflect the autocrine production of transforming growth factor-β1

1996 ◽  
Vol 148 (1) ◽  
pp. 87-94 ◽  
Author(s):  
A J Cowin ◽  
E L Heaton ◽  
S H Cheshire ◽  
S P Bidey
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Transforming Growth Factor ◽  
Thymidine Incorporation ◽  
Transforming Growth Factor Β1 ◽  
Follicular Cells ◽  
Stimulatory Action ◽  
Thyroid Follicular Cells ◽  
Epidermal Growth ◽  
Tgf Β1

Abstract The present study has investigated an involvement of autocrine transforming growth factor-β1 (TGF-β1) in regulating the proliferative response of porcine thyroid follicular cells (TFCs) to epidermal growth factor (EGF) and TSH. Primary monolayer TFC cultures exposed to EGF over the range 0–0·4 nmol/l showed a dose-dependent increase in [methyl-3H]thymidine incorporation, whereas higher EGF doses were associated with a reduction in the level of [methyl-3 H]thymidine incorporation. TGF-β immunoneutralisation had little effect on the stimulatory action of low EGF doses, but led to an increase in [methyl-3H]thymidine incorporation at higher EGF levels. In TFC cultures exposed to TSH, the level of [methyl-3H]thymidine incorporation attained at a dose of 1 U TSH/1 was enhanced in the presence of TGF-β1 antiserum, although the similar stimulatory effect of 8-bromo cAMP was unaffected. Treatment of TFCs with phorbol 12-myristate 13-acetate (8 nmol/l) to activate protein kinase C (PKC) led to an enhanced incorporation of [methyl-3H]thymidine which was increased further after neutralisation of endogenous TGF-β1. While confirming, therefore, a role for autocrine TGF-β1 in maintaining control of TFC DNA synthesis in vitro, these findings provide evidence that an increase in the availability of autocrine TGF-β1 effected by EGF and TSH may play an instrumental role in limiting the cellular hyperplasia induced by these factors within the thyroid follicular microenvironment. Moreover, the present data also suggest that the availability of active autocrine TGF-β1 to TFCs under such conditions may be dependent upon a PKC-mediated mechanism. Journal of Endocrinology (1996) 148, 87–94

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