臺灣博碩士論文加值系統

甲狀腺素(T3)可藉由與甲狀腺受體(thyroid hormone receptor, TR)的結合，活化TR及其相關的訊息途徑，以達到調控生長、發育、分化及代謝過程。TR和TR4 (testicular orphan receptor 4)均屬於細胞核內激素受體(nuclear hormone receptor)的家族成員，也是ligand-dependent轉錄因子。由於TR4與TR具有結合至相似的response element構形，即所謂中間包含四個核苷酸所構成的direct repeat，故本研究欲探討TR及TR4這兩條訊息途徑之間是否有cross-talk的可能。研究發現，TR經由不同甲狀腺素response elements (thyroid hormone response elements, TREs) 所調控的轉錄活性，會因為TR4的存在而有近92%受到抑制。而glutathione S-transferase (GST) pull-down assay是用來瞭解TR4抑制TR調控的轉錄活性，是否因為TR4與TR之間有交互作用所造成的。結果顯示，TRα1的C domain及TRβ1的A/B和C domain與TR4有交互作用；而TR4則是透過DNA和ligand binding domain與TRα1或TRβ1有交互作用。Electrophoretic mobility shift assay (EMSA)則是為了進一步瞭解TR4抑制T3所調控的訊息機制。實驗結果發現，隨著TR4的加入，TR homodimer或TR/RXR heterodimer的訊號會比沒有TR4存在時減少近65%。因此，EMSA和GST pull-down assay不僅證實TR4和TR之間有直接結合的關係，更說明TR4的存在對抑制TR所調控的轉錄活性是非常重要的。此外，在持續表現TRα1的肝癌細胞株中(HepG2-TRα1)，處理100 nM T3 48小時後，發現furin的蛋白質及mRNA表現量分別增加約3.4及2.8倍。然而，promoter assay卻發現，細胞處理10 nM T3 24小時後，furin起動子(promoter)的活性增加約5倍之多。不過，隨著TR4蛋白量的表現愈多，furin起動子的活性卻會減少約20至60%。這種抑制的情形，同樣在處理T3後，所造成的furin蛋白質及mRNA表現量上升，也因TR4的存在而受到抑制。另外，α-fetoprotein (AFP)是一種受到T3負向調控(negatively regulation)的基因，這種因T3的存在而抑制其表現的結果，可因同時存在TR4的情況下有回復的情形。綜合以上結果，TR和TR4之間的交互作用可能可以減少TR結合至同源TRE。意即TR4與TR之間的結合，可以改變因T3存在下，所造成的TR標的基因(target gene)的表現或抑制。研究結果認為，經由T3/TR所調控的furin及AFP的轉錄層次(transcription level)，可能會受TR4的影響。因此，本研究提出TRs和TR4之間存在cross-talk及可能有拮抗存在的機制。

Thyroid hormones (T3) regulate growth, development, differentiation, and metabolic processes by interacting with and activating thyroid hormone receptors or associated pathways. Both thyroid hormone receptor (TR) and testicular orphan receptor 4 (TR4) belong to the nuclear hormone receptor superfamily and are ligand-dependent transcription factors. Because of TR4 and TR sharing similar response element configurations, known as a direct repeat with four nucleotide spacings, this study investigated the possibility of cross-talk between the two ligand-dependent signal transduction pathways. The transcriptional activity of TR mediated via various thyroid hormone response elements (TREs) reporter construct was repressed by TR4 by up to 92%. The glutathione S-transferase (GST) pull-down assay was performed to determine whether the TR4 interacts with TR to account for the TR4 repressed the TR trans-activation. And the results indicate that both the C (DNA binding) domain of TRα1 and the A/B and C domain of TRβ1 interact with the TR4 protein while the TR4 interacts with the TRα1 or TRβ1 proteins through the DNA and ligand binding domains. To further understand the mechanism of TR4 suppresses T3-signalling, electrophoretic mobility shift assay (EMSA) was carried out. TR homodimer or TR/RXR (retinoid X receptor) heterodimer reduced up to 65% by adding increasing amounts of TR4 protein. Therefore, EMSA and GST pull-down assays demonstrated the direct binding of TR proteins to TR4 and revealed that the interaction is important to the TR4-mediated suppression of TR-transactivation. Additionally, furin was elevated approximately 3.4-fold and 2.8-fold in HepG2-TRα1 cells at the protein and mRNA levels, respectively, after 48 hr of 100 nM T3 treatment. Increasing expression of TR4 repressed approximately 20 to 60% of furin promoter activities, which were stimulated roughly 5-fold by 10 nM T3 for 24 hr. Similarly, the up-regulated furin protein and mRNA levels by T3 were gradually suppressed by increasing amounts of TR4. In addition, α-fetoprotein (AFP) is negatively regulated by T3. And the suppression of AFP by T3 was gradually reversed by co-transfection with increasing amounts of TR4 expression plasmid. Taken together, interaction of TR4 with TR proteins may reduce TR binding to its cognate TRE. Subsequently, the interaction between TR4 and TR alters TR target gene stimulation or repression by T3. These findings suggest that TR4 may influence furin and AFP metabolism regulated by T3/TR at the transcriptional level. This study proposes a mechanism for cross-talk and potential antagonism between TRs and TR4.

指導教授推薦書…………………………………………………………………I
口試委員審定書………………………………………………………………..II
授權書…………………………………………………………………………III
誌謝……………………………………………………………………………...VI
Abbreviations…………………………………………………………………VII
摘要……………………………………………………………………………IX
Abstract………………………………………………………………………...XI
Introduction……………………………...……………………………………...1
Nuclear receptor superfamily………………………………………………….1
Orphan receptor………………………………………………………………..3
Testicular orphan receptor 4 (TR4)…………………………………………..3
Function of TR4………………………………………………………………...4
Thyroid hormone receptor (TR)………………………………………………5
Thyroid hormone receptor isoforms…………………………………………..6
Thyroid hormone response elements………………………………………….7
Thyroid hormone action in mouse models……………………………………8
Liver is a target organ for TRs……………………………………………….10
Furin– T3 up-regulated gene…………………………………………………11
α-Fetoprotein– T3 down-regulated gene……………………………………..12
Materials and Methods………………………………………………………13
Plasmids……………………………………………………………………….13
Cell culture…………………………………………………………………….13
Immunoblot analysis………………………………………………………….14
Northern blot analysis………………………………………………………...15
Quantitative reverse transcription polymerase chain reaction (Q-RT-PCR)..
.............................................................................................................................15
GST pull-down assay…………………………………………………………16
Determination of the trans-activation activity of TRs……………………...17
Electrophoretic mobility-shift assay (EMSA)……………………………….18
Results…………………………………………………………………………...19
Suppression of T3-dependent transactivation activity by TR4……………19
Interaction of TR4 with TRs…………………………………………………20
Interruption of TR/TR or TR/RXR binding via the DNA and ligand binding domains of TR4………………………………………………………………..22
Effects of T3 on the abundance of furin protein and mRNA in HepG2-TRα1 cells......................................................................................................................23
Suppression of TR-target genes expression by TR4………………………...25
Discussion……………………………………………………………………….27
Figures…………………………………………………………………………33
Fig. 1. Structure/function organization of nuclear receptors………………33
Fig. 2. Molecular mechanism of nuclear receptor action…………………...34
Fig. 3. General model for thyroid hormone action in the nucleus…………35
Fig. 4. Comparison of amino acid homologies and their functional pro- perties among TR isoforms…………………………………………………...36
Fig. 5. Half-site orientation and optimal nucleotide spacing between half- sites……………………………………………………………………………..37
Fig. 6. Repression of T3-dependent transcriptional activity in CV1, HEK293 and HepG2-TRα1 cells by TR4………………………………………………38
Fig. 7. Interaction between TRs and TR4…………………………………...40
Fig. 8. TRβ1–ABC domains interact with TR4……………………………...42
Fig. 9. TRα1–C domains interact with TR4…………………………………44
Fig. 10. TR4–CDE domains interact with TRα1 and TRβ1………………..46
Fig. 11. Binding of TR and TR4 proteins to F2-TRE by EMSA…………...48
Fig. 12. Effect of T3 on furin protein expression in HepG2-TRα1 cells…………......................................................................................................50
Fig. 13. Effect of T3 on the amount of furin mRNA in HepG2 TRα1 cell lines….................................................................................................................51
Fig. 14. Repression of T3-target gene furin by TR4………………………...52
Fig. 15. Binding of TR and TR4 proteins to furin-TRE by EMSA………54
Fig. 16. Relief of T3 suppressive effect on α-fetoprotein by TR4…………...55
References………………………………………………………………………57

1.Bourguet W, Germain P, Gronemeyer H 2000 Nuclear receptor ligand-binding domains: three-dimensional structures, molecular interactions and pharmacological implications. Trends Pharmacol Sci 21:381-8
2.Mangelsdorf DJ, Evans RM 1995 The RXR heterodimers and orphan receptors. Cell 83:841-50
3.Olefsky JM 2001 Nuclear receptor minireview series. J Biol Chem 276:36863-4
4.Mangelsdorf DJ, Thummel C, Beato M, et al. 1995 The nuclear receptor superfamily: the second decade. Cell 83:835-9
5.Tsai MJ, O'Malley BW 1994 Molecular mechanisms of action of steroid/thyroid receptor superfamily members. Annu Rev Biochem 63:451-86
6.O'Malley BW, Tsai MJ 1992 Molecular pathways of steroid receptor action. Biol Reprod 46:163-7
7.Truss M, Beato M 1993 Steroid hormone receptors: interaction with deoxyribonucleic acid and transcription factors. Endocr Rev 14:459-79
8.Lee YF, Lee HJ, Chang C 2002 Recent advances in the TR2 and TR4 orphan receptors of the nuclear receptor superfamily. J Steroid Biochem Mol Biol 81:291-308
9.Lee H, Yen PM 1999 Recent advances in understanding thyroid hormone receptor coregulators. J Biomed Sci 6:71-8
10.Laudet V 1997 Evolution of the nuclear receptor superfamily: early diversification from an ancestral orphan receptor. J Mol Endocrinol 19:207-26
11.Chang C, Da Silva SL, Ideta R, Lee Y, Yeh S, Burbach JP 1994 Human and rat TR4 orphan receptors specify a subclass of the steroid receptor superfamily. Proc Natl Acad Sci U S A 91:6040-4
12.Hirose T, Fujimoto W, Tamaai T, Kim KH, Matsuura H, Jetten AM 1994 TAK1: molecular cloning and characterization of a new member of the nuclear receptor superfamily. Mol Endocrinol 8:1667-80
13.Yoshikawa T, DuPont BR, Leach RJ, Detera-Wadleigh SD 1996 New variants of the human and rat nuclear hormone receptor, TR4: expression and chromosomal localization of the human gene. Genomics 35:361-6
14.Lee HJ, Lee Y, Burbach JP, Chang C 1995 Suppression of gene expression on the simian virus 40 major late promoter by human TR4 orphan receptor. A member of the steroid receptor superfamily. J Biol Chem 270:30129-33
15.Lee YF, Pan HJ, Burbach JP, Morkin E, Chang C 1997 Identification of direct repeat 4 as a positive regulatory element for the human TR4 orphan receptor. A modulator for the thyroid hormone target genes. J Biol Chem 272:12215-20
16.Lee YF, Young WJ, Burbach JP, Chang C 1998 Negative feedback control of the retinoid-retinoic acid/retinoid X receptor pathway by the human TR4 orphan receptor, a member of the steroid receptor superfamily. J Biol Chem 273:13437-43
17.Lee YF, Young WJ, Lin WJ, Shyr CR, Chang C 1999 Differential regulation of direct repeat 3 vitamin D3 and direct repeat 4 thyroid hormone signaling pathways by the human TR4 orphan receptor. J Biol Chem 274:16198-205
18.Young WJ, Smith SM, Chang C 1997 Induction of the intronic enhancer of the human ciliary neurotrophic factor receptor (CNTFRalpha) gene by the TR4 orphan receptor. A member of steroid receptor superfamily. J Biol Chem 272:3109-16
19.Shyr CR, Hu YC, Kim E, Chang C 2002 Modulation of estrogen receptor-mediated transactivation by orphan receptor TR4 in MCF-7 cells. J Biol Chem 277:14622-8
20.Yan ZH, Karam WG, Staudinger JL, Medvedev A, Ghanayem BI, Jetten AM 1998 Regulation of peroxisome proliferator-activated receptor alpha-induced transactivation by the nuclear orphan receptor TAK1/TR4. J Biol Chem 273:10948-57
21.Lee YF, Shyr CR, Thin TH, Lin WJ, Chang C 1999 Convergence of two repressors through heterodimer formation of androgen receptor and testicular orphan receptor-4: a unique signaling pathway in the steroid receptor superfamily. Proc Natl Acad Sci U S A 96:14724-9
22.Cheng SY 2000 Multiple mechanisms for regulation of the transcriptional activity of thyroid hormone receptors. Rev Endocr Metab Disord 1:9-18
23.Oppenheimer JH, Schwartz HL, Mariash CN, Kinlaw WB, Wong NC, Freake HC 1987 Advances in our understanding of thyroid hormone action at the cellular level. Endocr Rev 8:288-308
24.Samuels HH, Forman BM, Horowitz ZD, Ye ZS 1988 Regulation of gene expression by thyroid hormone. J Clin Invest 81:957-67
25.Yen PM 2001 Physiological and molecular basis of thyroid hormone action. Physiol Rev 81:1097-142
26.Sap J, Munoz A, Damm K, et al. 1986 The c-erb-A protein is a high-affinity receptor for thyroid hormone. Nature 324:635-40
27.Weinberger C, Thompson CC, Ong ES, Lebo R, Gruol DJ, Evans RM 1986 The c-erb-A gene encodes a thyroid hormone receptor. Nature 324:641-6
28.Loosfelt H, Atger M, Misrahi M, et al. 1986 Cloning and sequence analysis of rabbit progesterone-receptor complementary DNA. Proc Natl Acad Sci U S A 83:9045-9
29.Cheng Sy S 1995 New Insights into the Structure and Function of the Thyroid Hormone Receptor. J Biomed Sci 2:77-89
30.Munoz A, Zenke M, Gehring U, Sap J, Beug H, Vennstrom B 1988 Characterization of the hormone-binding domain of the chicken c-erbA/thyroid hormone receptor protein. Embo J 7:155-9
31.Forman BM, Yang CR, Stanley F, Casanova J, Samuels HH 1988 c-erbA protooncogenes mediate thyroid hormone-dependent and independent regulation of the rat growth hormone and prolactin genes. Mol Endocrinol 2:902-11
32.Schueler PA, Schwartz HL, Strait KA, Mariash CN, Oppenheimer JH 1990 Binding of 3,5,3'-triiodothyronine (T3) and its analogs to the in vitro translational products of c-erbA protooncogenes: differences in the affinity of the alpha- and beta-forms for the acetic acid analog and failure of the human testis and kidney alpha-2 products to bind T3. Mol Endocrinol 4:227-34
33.Hodin RA, Lazar MA, Chin WW 1990 Differential and tissue-specific regulation of the multiple rat c-erbA messenger RNA species by thyroid hormone. J Clin Invest 85:101-5
34.Bradley DJ, Towle HC, Young WS, 3rd 1992 Spatial and temporal expression of alpha- and beta-thyroid hormone receptor mRNAs, including the beta 2-subtype, in the developing mammalian nervous system. J Neurosci 12:2288-302
35.Bradley DJ, Towle HC, Young WS, 3rd 1994 Alpha and beta thyroid hormone receptor (TR) gene expression during auditory neurogenesis: evidence for TR isoform-specific transcriptional regulation in vivo. Proc Natl Acad Sci U S A 91:439-43
36.Glass CK 1994 Differential recognition of target genes by nuclear receptor monomers, dimers, and heterodimers. Endocr Rev 15:391-407
37.Bigler J, Eisenman RN 1995 Novel location and function of a thyroid hormone response element. Embo J 14:5710-23
38.Zhang W, Brooks RL, Silversides DW, et al. 1992 Negative thyroid hormone control of human growth hormone gene expression is mediated by 3'-untranslated/3'-flanking DNA. J Biol Chem 267:15056-63
39.Onate SA, Tsai SY, Tsai MJ, O'Malley BW 1995 Sequence and characterization of a coactivator for the steroid hormone receptor superfamily. Science 270:1354-7
40.Chen JD, Evans RM 1995 A transcriptional co-repressor that interacts with nuclear hormone receptors. Nature 377:454-7
41.Yu F, Gothe S, Wikstrom L, Forrest D, Vennstrom B, Larsson L 2000 Effects of thyroid hormone receptor gene disruption on myosin isoform expression in mouse skeletal muscles. Am J Physiol Regul Integr Comp Physiol 278:R1545-54
42.Johansson C, Lannergren J, Lunde PK, Vennstrom B, Thoren P, Westerblad H 2000 Isometric force and endurance in soleus muscle of thyroid hormone receptor-alpha(1)- or -beta-deficient mice. Am J Physiol Regul Integr Comp Physiol 278:R598-603
43.Dellovade TL, Chan J, Vennstrom B, Forrest D, Pfaff DW 2000 The two thyroid hormone receptor genes have opposite effects on estrogen-stimulated sex behaviors. Nat Neurosci 3:472-5
44.Abel ED, Boers ME, Pazos-Moura C, et al. 1999 Divergent roles for thyroid hormone receptor beta isoforms in the endocrine axis and auditory system. J Clin Invest 104:291-300
45.Johansson C, Gothe S, Forrest D, Vennstrom B, Thoren P 1999 Cardiovascular phenotype and temperature control in mice lacking thyroid hormone receptor-beta or both alpha1 and beta. Am J Physiol 276:H2006-12
46.Chamba A, Neuberger J, Strain A, Hopkins J, Sheppard MC, Franklyn JA 1996 Expression and function of thyroid hormone receptor variants in normal and chronically diseased human liver. J Clin Endocrinol Metab 81:360-7
47.Lin KH, Shieh HY, Hsu HC 2000 Negative regulation of the antimetastatic gene Nm23-H1 by thyroid hormone receptors. Endocrinology 141:2540-7
48.Seidah NG, Day R, Marcinkiewicz M, Chretien M 1998 Precursor convertases: an evolutionary ancient, cell-specific, combinatorial mechanism yielding diverse bioactive peptides and proteins. Ann N Y Acad Sci 839:9-24
49.Thacker C, Rose AM 2000 A look at the Caenorhabditis elegans Kex2/Subtilisin-like proprotein convertase family. Bioessays 22:545-53
50.Zhou A, Webb G, Zhu X, Steiner DF 1999 Proteolytic processing in the secretory pathway. J Biol Chem 274:20745-8
51.Seidah NG, Chretien M 1999 Proprotein and prohormone convertases: a family of subtilases generating diverse bioactive polypeptides. Brain Res 848:45-62
52.2002 From the Centers for Disease Control and Prevention. Update: investigation of bioterrorism-related Anthrax--Connecticut, 2001. Jama 287:34-5
53.Steiner DF 1998 The proprotein convertases. Curr Opin Chem Biol 2:31-9
54.van den Ouweland AM, van Duijnhoven HL, Keizer GD, Dorssers LC, Van de Ven WJ 1990 Structural homology between the human fur gene product and the subtilisin-like protease encoded by yeast KEX2. Nucleic Acids Res 18:664
55.Bravo DA, Gleason JB, Sanchez RI, Roth RA, Fuller RS 1994 Accurate and efficient cleavage of the human insulin proreceptor by the human proprotein-processing protease furin. Characterization and kinetic parameters using the purified, secreted soluble protease expressed by a recombinant baculovirus. J Biol Chem 269:25830-7
56.Jean F, Stella K, Thomas L, et al. 1998 alpha1-Antitrypsin Portland, a bioengineered serpin highly selective for furin: application as an antipathogenic agent. Proc Natl Acad Sci U S A 95:7293-8
57.Cameron A, Appel J, Houghten RA, Lindberg I 2000 Polyarginines are potent furin inhibitors. J Biol Chem 275:36741-9
58.van de Ven WJ, Voorberg J, Fontijn R, et al. 1990 Furin is a subtilisin-like proprotein processing enzyme in higher eukaryotes. Mol Biol Rep 14:265-75
59.Anderson ED, VanSlyke JK, Thulin CD, Jean F, Thomas G 1997 Activation of the furin endoprotease is a multiple-step process: requirements for acidification and internal propeptide cleavage. Embo J 16:1508-18
60.Thomas G 2002 Furin at the cutting edge: from protein traffic to embryogenesis and disease. Nat Rev Mol Cell Biol 3:753-66
61.Sell S, Becker FF 1978 alpha-Fetoprotein. J Natl Cancer Inst 60:19-26
62.Van Reeth T, Gabant P, Szpirer C, Szpirer J 2002 Stimulation of the alpha-fetoprotein promoter by unliganded thyroid hormone receptor in association with protein deacetylation. Mol Cell Endocrinol 188:99-109
63.Caturla M, Van Reeth T, Dreze P, Szpirer J, Szpirer C 1997 The thyroid hormone down-regulates the mouse alpha-foetoprotein promoter. Mol Cell Endocrinol 135:139-45
64.Toublanc JE, Ingrand J, Megarbane M, Fulla Y, Rives S 1994 [Alpha fetoprotein in congenital hypothyroidism before and during treatment]. Arch Pediatr 1:872-8
65.Samuels HH, Stanley F, Casanova J 1979 Depletion of L-3,5,3'-triiodothyronine and L-thyroxine in euthyroid calf serum for use in cell culture studies of the action of thyroid hormone. Endocrinology 105:80-5
66.Bhat MK, McPhie P, Ting YT, Zhu XG, Cheng SY 1995 Structure of the carboxy-terminal region of thyroid hormone nuclear receptors and its possible role in hormone-dependent intermolecular interactions. Biochemistry 34:10591-9
67.Lin KH, Wang WJ, Wu YH, Cheng SY 2002 Activation of antimetastatic Nm23-H1 gene expression by estrogen and its alpha-receptor. Endocrinology 143:467-75
68.Shih CH, Chen SL, Yen CC, et al. 2004 Thyroid hormone receptor-dependent transcriptional regulation of fibrinogen and coagulation proteins. Endocrinology 145:2804-14
69.Lin KH, Chen CY, Chen SL, et al. 2004 Regulation of fibronectin by thyroid hormone receptors. J Mol Endocrinol 33:445-58
70.Kyriakis JM, Avruch J 1996 Protein kinase cascades activated by stress and inflammatory cytokines. Bioessays 18:567-77
71.Lin KH, Zhu XG, Hsu HC, et al. 1997 Dominant negative activity of mutant thyroid hormone alpha1 receptors from patients with hepatocellular carcinoma. Endocrinology 138:5308-15
72.Flores-Morales A, Gullberg H, Fernandez L, et al. 2002 Patterns of liver gene expression governed by TRbeta. Mol Endocrinol 16:1257-68
73.Schmilovitz-Weiss H, Stemmer SM, Liberzon E, et al. 2006 Quantitation of alpha-fetoprotein messenger RNA for early detection of recurrent hepatocellular carcinoma: A prospective pilot study. Cancer Detect Prev 30:204-9
74.Ayoubi TA, Creemers JW, Roebroek AJ, Van de Ven WJ 1994 Expression of the dibasic proprotein processing enzyme furin is directed by multiple promoters. J Biol Chem 269:9298-303
75.Franco PJ, Farooqui M, Seto E, Wei LN 2001 The orphan nuclear receptor TR2 interacts directly with both class I and class II histone deacetylases. Mol Endocrinol 15:1318-28
76.Hwang SB, Burbach JP, Chang C 1998 TR4 orphan receptor crosstalks to chicken ovalbumin upstream protein-transcription factor and thyroid hormone receptor to induce the transcriptional activity of the human immunodeficiency virus type 1 long-terminal repeat. Endocrine 8:169-75
77.Lee HJ, Lee YF, Chang C 2001 TR4 orphan receptor represses the human steroid 21-hydroxylase gene expression through the monomeric AGGTCA motif. Biochem Biophys Res Commun 285:1361-8
78.Lin WJ, Li J, Lee YF, et al. 2003 Suppression of hepatitis B virus core promoter by the nuclear orphan receptor TR4. J Biol Chem 278:9353-60
79.Collins LL, Lee YF, Heinlein CA, et al. 2004 Growth retardation and abnormal maternal behavior in mice lacking testicular orphan nuclear receptor 4. Proc Natl Acad Sci U S A 101:15058-63
80.Yaffe BM, Samuels HH 1984 Hormonal regulation of the growth hormone gene. Relationship of the rate of transcription to the level of nuclear thyroid hormone-receptor complexes. J Biol Chem 259:6284-91
81.Palomino T, Barettino D, Aranda A 1998 Role of GHF-1 in the regulation of the rat growth hormone gene promoter by thyroid hormone and retinoic acid receptors. J Biol Chem 273:27541-7
82.Gevers EF, van der Eerden BC, Karperien M, Raap AK, Robinson IC, Wit JM 2002 Localization and regulation of the growth hormone receptor and growth hormone-binding protein in the rat growth plate. J Bone Miner Res 17:1408-19
83.Raber W, Nowotny P, Vytiska-Binstorfer E, Vierhapper H 2003 Thyroxine treatment modified in infertile women according to thyroxine-releasing hormone testing: 5 year follow-up of 283 women referred after exclusion of absolute causes of infertility. Hum Reprod 18:707-14
84.Jiang JY, Imai Y, Umezu M, Sato E 2001 Characteristics of infertility in female hypothyroid (hyt) mice. Reproduction 122:695-700
85.Johnson C, Olivier NB, Nachreiner R, Mullaney T 1999 Effect of 131I-induced hypothyroidism on indices of reproductive function in adult male dogs. J Vet Intern Med 13:104-10
86.Tahmaz L, Gokalp A, Kibar Y, Kocak I, Yalcin O, Ozercan Y 2000 Effect of hypothyroidism on the testes in mature rats and treatment with levothyroxine and zinc. Andrologia 32:85-9
87.Lombardo F, Sgro P, Salacone P, et al. 2005 Androgens and fertility. J Endocrinol Invest 28:51-5
88.Mu X, Lee YF, Liu NC, et al. 2004 Targeted inactivation of testicular nuclear orphan receptor 4 delays and disrupts late meiotic prophase and subsequent meiotic divisions of spermatogenesis. Mol Cell Biol 24:5887-99
89.Kim E, Yang Z, Liu NC, Chang C 2005 Induction of apolipoprotein E expression by TR4 orphan nuclear receptor via 5' proximal promoter region. Biochem Biophys Res Commun 328:85-90
90.Kim E, Xie S, Yeh SD, et al. 2003 Disruption of TR4 orphan nuclear receptor reduces the expression of liver apolipoprotein E/C-I/C-II gene cluster. J Biol Chem 278:46919-26
91.Vandenbrouck Y, Janvier B, Loriette C, Bereziat G, Mangeney-Andreani M 1994 The modulation of apolipoprotein E gene expression by 3,3'-5-triiodothyronine in HepG2 cells occurs at transcriptional and post-transcriptional levels. Eur J Biochem 224:463-71
92.Bassi DE, Mahloogi H, Lopez De Cicco R, Klein-Szanto A 2003 Increased furin activity enhances the malignant phenotype of human head and neck cancer cells. Am J Pathol 162:439-47
93.Siegfried G, Basak A, Cromlish JA, et al. 2003 The secretory proprotein convertases furin, PC5, and PC7 activate VEGF-C to induce tumorigenesis. J Clin Invest 111:1723-32
94.Bassi DE, Mahloogi H, Klein-Szanto AJ 2000 The proprotein convertases furin and PACE4 play a significant role in tumor progression. Mol Carcinog 28:63-9
95.Cheng M, Watson PH, Paterson JA, Seidah N, Chretien M, Shiu RP 1997 Pro-protein convertase gene expression in human breast cancer. Int J Cancer 71:966-71
96.McMahon S, Laprise MH, Dubois CM 2003 Alternative pathway for the role of furin in tumor cell invasion process. Enhanced MMP-2 levels through bioactive TGFbeta. Exp Cell Res 291:326-39