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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 鄭永銘(Yung-Ming Jeng) | |
dc.contributor.author | Yu-Lin Jhuang | en |
dc.contributor.author | 莊郁琳 | zh_TW |
dc.date.accessioned | 2021-06-16T06:45:26Z | - |
dc.date.available | 2019-10-09 | |
dc.date.copyright | 2014-10-09 | |
dc.date.issued | 2014 | |
dc.date.submitted | 2014-07-28 | |
dc.identifier.citation | 1. Sorlie T, Perou CM, Tibshirani R, et al., Gene expression patterns of breast carcinomas distinguish tumor subclasses with clinical implications. Proc Natl Acad Sci U S A, 2001;19:10869-74.
2. Sarrio D, Rodriguez-Pinilla SM, Hardisson D, et al., Epithelial-mesenchymal transition in breast cancer relates to the basal-like phenotype. Cancer Res, 2008;4:989-97. 3. Rakha EA, Reis-Filho JS, and Ellis IO, Basal-like breast cancer: a critical review. J Clin Oncol, 2008;15:2568-81. 4. Carey L, Winer E, Viale G, et al., Triple-negative breast cancer: disease entity or title of convenience? Nat Rev Clin Oncol, 2010;12:683-92. 5. Hay ED, An overview of epithelio-mesenchymal transformation. Acta Anat (Basel), 1995;1:8-20. 6. Proud D and Leigh R, Epithelial cells and airway diseases. Immunol Rev, 2011;1:186-204. 7. Artis D, Epithelial-cell recognition of commensal bacteria and maintenance of immune homeostasis in the gut. Nat Rev Immunol, 2008;6:411-20. 8. Lamouille S, Xu J, and Derynck R, Molecular mechanisms of epithelial-mesenchymal transition. Nat Rev Mol Cell Biol, 2014;3:178-96. 9. Kalluri R and Neilson EG, Epithelial-mesenchymal transition and its implications for fibrosis. Journal of Clinical Investigation, 2003;12:1776-1784. 10. Kalluri R and Weinberg RA, The basics of epithelial-mesenchymal transition. J Clin Invest, 2009;6:1420-8. 11. Strutz F, Okada H, Lo CW, et al., Identification and characterization of a fibroblast marker: FSP1. J Cell Biol, 1995;2:393-405. 12. Mani SA, Guo W, Liao MJ, et al., The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell, 2008;4:704-15. 13. Thompson EW, Newgreen DF, and Tarin D, Carcinoma invasion and metastasis: a role for epithelial-mesenchymal transition? Cancer Res, 2005;14:5991-5; discussion 5995. 14. Zeisberg M and Neilson EG, Biomarkers for epithelial-mesenchymal transitions. J Clin Invest, 2009;6:1429-37. 15. Thiery JP, Epithelial-mesenchymal transitions in tumour progression. Nat Rev Cancer, 2002;6:442-54. 16. Birchmeier W and Behrens J, Cadherin expression in carcinomas: role in the formation of cell junctions and the prevention of invasiveness. Biochim Biophys Acta, 1994;1:11-26. 17. Behrens J, Mareel MM, Van Roy FM, et al., Dissecting tumor cell invasion: epithelial cells acquire invasive properties after the loss of uvomorulin-mediated cell-cell adhesion. J Cell Biol, 1989;6:2435-47. 18. Moody SE, Perez D, Pan TC, et al., The transcriptional repressor Snail promotes mammary tumor recurrence. Cancer Cell, 2005;3:197-209. 19. Ponzo MG and Park M, The Met receptor tyrosine kinase and basal breast cancer. Cell Cycle, 2010;6:1043-50. 20. Di Renzo MF, Olivero M, Ferro S, et al., Overexpression of the c-MET/HGF receptor gene in human thyroid carcinomas. Oncogene, 1992;12:2549-53. 21. Di Renzo MF, Poulsom R, Olivero M, et al., Expression of the Met/hepatocyte growth factor receptor in human pancreatic cancer. Cancer Res, 1995;5:1129-38. 22. Di Renzo MF, Olivero M, Giacomini A, et al., Overexpression and amplification of the met/HGF receptor gene during the progression of colorectal cancer. Clin Cancer Res, 1995;2:147-54. 23. Massague J, TGF-beta signal transduction. Annu Rev Biochem, 1998:753-91. 24. Herpin A, Lelong C, and Favrel P, Transforming growth factor-beta-related proteins: an ancestral and widespread superfamily of cytokines in metazoans. Dev Comp Immunol, 2004;5:461-85. 25. Derynck R, Akhurst RJ, and Balmain A, TGF-beta signaling in tumor suppression and cancer progression. Nat Genet, 2001;2:117-29. 26. Miettinen PJ, Ebner R, Lopez AR, et al., TGF-beta induced transdifferentiation of mammary epithelial cells to mesenchymal cells: involvement of type I receptors. J Cell Biol, 1994;6 Pt 2:2021-36. 27. Massague J, TGFbeta in Cancer. Cell, 2008;2:215-30. 28. Sun PD and Davies DR, The cystine-knot growth-factor superfamily. Annu Rev Biophys Biomol Struct, 1995:269-91. 29. Wrana JL, Attisano L, Wieser R, et al., Mechanism of activation of the TGF-beta receptor. Nature, 1994;6488:341-7. 30. Wieser R, Wrana JL, and Massague J, GS domain mutations that constitutively activate T beta R-I, the downstream signaling component in the TGF-beta receptor complex. EMBO J, 1995;10:2199-208. 31. McPherron AC, Lawler AM, and Lee SJ, Regulation of skeletal muscle mass in mice by a new TGF-beta superfamily member. Nature, 1997;6628:83-90. 32. Derynck R and Zhang YE, Smad-dependent and Smad-independent pathways in TGF-beta family signalling. Nature, 2003;6958:577-84. 33. Moustakas A and Heldin CH, Non-Smad TGF-beta signals. J Cell Sci, 2005;Pt 16:3573-84. 34. Xu J, Lamouille S, and Derynck R, TGF-beta-induced epithelial to mesenchymal transition. Cell Res, 2009;2:156-72. 35. Horiguchi K, Sakamoto K, Koinuma D, et al., TGF-beta drives epithelial-mesenchymal transition through deltaEF1-mediated downregulation of ESRP. Oncogene, 2012;26:3190-201. 36. Wu CY, Tsai YP, Wu MZ, et al., Epigenetic reprogramming and post-transcriptional regulation during the epithelial-mesenchymal transition. Trends Genet, 2012;9:454-63. 37. Lin T, Ponn A, Hu X, et al., Requirement of the histone demethylase LSD1 in Snai1-mediated transcriptional repression during epithelial-mesenchymal transition. Oncogene, 2010;35:4896-904. 38. Yi JY, Shin I, and Arteaga CL, Type I transforming growth factor beta receptor binds to and activates phosphatidylinositol 3-kinase. J Biol Chem, 2005;11:10870-6. 39. Sarbassov DD, Ali SM, and Sabatini DM, Growing roles for the mTOR pathway. Curr Opin Cell Biol, 2005;6:596-603. 40. Lamouille S and Derynck R, Cell size and invasion in TGF-beta-induced epithelial to mesenchymal transition is regulated by activation of the mTOR pathway. J Cell Biol, 2007;3:437-51. 41. Bakin AV, Tomlinson AK, Bhowmick NA, et al., Phosphatidylinositol 3-kinase function is required for transforming growth factor beta-mediated epithelial to mesenchymal transition and cell migration. J Biol Chem, 2000;47:36803-10. 42. Lehmann K, Raf induces TGFbeta production while blocking its apoptotic but not invasive responses: a mechanism leading to increased malignancy in epithelial cells. Genes & Development, 2000;20:2610-2622. 43. Janda E, Lehmann K, Killisch I, et al., Ras and TGF[beta] cooperatively regulate epithelial cell plasticity and metastasis: dissection of Ras signaling pathways. J Cell Biol, 2002;2:299-313. 44. Egeblad M and Werb Z, New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer, 2002;3:161-74. 45. Mott JD and Werb Z, Regulation of matrix biology by matrix metalloproteinases. Curr Opin Cell Biol, 2004;5:558-64. 46. Duivenvoorden WC, Hirte HW, and Singh G, Transforming growth factor beta1 acts as an inducer of matrix metalloproteinase expression and activity in human bone-metastasizing cancer cells. Clin Exp Metastasis, 1999;1:27-34. 47. Kim ES, Sohn YW, and Moon A, TGF-beta-induced transcriptional activation of MMP-2 is mediated by activating transcription factor (ATF)2 in human breast epithelial cells. Cancer Lett, 2007;1:147-56. 48. Lin SW, Ke FC, Hsiao PW, et al., Critical involvement of ILK in TGFbeta1-stimulated invasion/migration of human ovarian cancer cells is associated with urokinase plasminogen activator system. Exp Cell Res, 2007;3:602-13. 49. Harbeck N, Kates RE, Gauger K, et al., Urokinase-type plasminogen activator (uPA) and its inhibitor PAI-I: novel tumor-derived factors with a high prognostic and predictive impact in breast cancer. Thromb Haemost, 2004;3:450-6. 50. Thali M, Muller MM, DeLorenzi M, et al., Drosophila homoeotic genes encode transcriptional activators similar to mammalian OTF-2. Nature, 1988;6199:598-601. 51. Stelnicki EJ, Arbeit J, Cass DL, et al., Modulation of the human homeobox genes PRX-2 and HOXB13 in scarless fetal wounds. J Invest Dermatol, 1998;1:57-63. 52. White P, Thomas DW, Fong S, et al., Deletion of the homeobox gene PRX-2 affects fetal but not adult fibroblast wound healing responses. J Invest Dermatol, 2003;1:135-44. 53. Meijlink F, Beverdam A, Brouwer A, et al., Vertebrate aristaless-related genes. Int J Dev Biol, 1999;7:651-63. 54. Lu MF, Cheng HT, Lacy AR, et al., Paired-related homeobox genes cooperate in handplate and hindlimb zeugopod morphogenesis. Dev Biol, 1999;1:145-57. 55. Doufexi AE and Mina M, Signaling pathways regulating the expression of Prx1 and Prx2 in the chick mandibular mesenchyme. Dev Dyn, 2008;11:3115-27. 56. Balic A, Adams D, and Mina M, Prx1 and Prx2 cooperatively regulate the morphogenesis of the medial region of the mandibular process. Dev Dyn, 2009;10:2599-613. 57. Debnath J, Muthuswamy SK, and Brugge JS, Morphogenesis and oncogenesis of MCF-10A mammary epithelial acini grown in three-dimensional basement membrane cultures. Methods, 2003;3:256-268. 58. Ocana OH, Corcoles R, Fabra A, et al., Metastatic colonization requires the repression of the epithelial-mesenchymal transition inducer Prrx1. Cancer Cell, 2012;6:709-24. 59. Huang FI, Chen YL, Chang CN, et al., Hepatocyte growth factor activates Wnt pathway by transcriptional activation of LEF1 to facilitate tumor invasion. Carcinogenesis, 2012;6:1142-8. 60. Vassalli JD, Sappino AP, and Belin D, The plasminogen activator/plasmin system. J Clin Invest, 1991;4:1067-72. 61. Broos S, Hulpiau P, Galle J, et al., ConTra v2: a tool to identify transcription factor binding sites across species, update 2011. Nucleic Acids Res, 2011;Web Server issue:W74-8. 62. Gupta GP and Massague J, Cancer metastasis: building a framework. Cell, 2006;4:679-95. 63. Peinado H, Quintanilla M, and Cano A, Transforming growth factor beta-1 induces snail transcription factor in epithelial cell lines: mechanisms for epithelial mesenchymal transitions. J Biol Chem, 2003;23:21113-23. 64. Morita T, Mayanagi T, and Sobue K, Dual roles of myocardin-related transcription factors in epithelial mesenchymal transition via slug induction and actin remodeling. J Cell Biol, 2007;5:1027-42. 65. Postigo AA, Opposing functions of ZEB proteins in the regulation of the TGFbeta/BMP signaling pathway. EMBO J, 2003;10:2443-52. 66. Sullivan NJ, Sasser AK, Axel AE, et al., Interleukin-6 induces an epithelial-mesenchymal transition phenotype in human breast cancer cells. Oncogene, 2009;33:2940-7. 67. Grotegut S, von Schweinitz D, Christofori G, et al., Hepatocyte growth factor induces cell scattering through MAPK/Egr-1-mediated upregulation of Snail. EMBO J, 2006;15:3534-45. 68. Billottet C, Tuefferd M, Gentien D, et al., Modulation of several waves of gene expression during FGF-1 induced epithelial-mesenchymal transition of carcinoma cells. J Cell Biochem, 2008;3:826-39. 69. Xie L, Law BK, Aakre ME, et al., Transforming growth factor beta-regulated gene expression in a mouse mammary gland epithelial cell line. Breast Cancer Res, 2003;6:R187-98. 70. Norris RA and Kern MJ, The identification of Prx1 transcription regulatory domains provides a mechanism for unequal compensation by the Prx1 and Prx2 loci. J Biol Chem, 2001;29:26829-37. 71. Takahashi Y, Sawada G, Kurashige J, et al., Paired related homoeobox 1, a new EMT inducer, is involved in metastasis and poor prognosis in colorectal cancer. Br J Cancer, 2013;2:307-11. 72. Poste G and Fidler IJ, The pathogenesis of cancer metastasis. Nature, 1980;5743:139-46. 73. Fidler IJ, The pathogenesis of cancer metastasis: the 'seed and soil' hypothesis revisited. Nat Rev Cancer, 2003;6:453-8. 74. Nakanishi K, Sakamoto M, Yasuda J, et al., Critical involvement of the phosphatidylinositol 3-kinase/Akt pathway in anchorage-independent growth and hematogeneous intrahepatic metastasis of liver cancer. Cancer Res, 2002;10:2971-5. 75. Gervais C, Mauvieux L, Perrusson N, et al., A new translocation t(9;11)(q34;p15) fuses NUP98 to a novel homeobox partner gene, PRRX2, in a therapy-related acute myeloid leukemia. Leukemia, 2005;1:145-8. 76. Lam DH and Aplan PD, NUP98 gene fusions in hematologic malignancies. Leukemia, 2001;11:1689-95. 77. Nakamura T, Yamazaki Y, Hatano Y, et al., NUP98 is fused to PMX1 homeobox gene in human acute myelogenous leukemia with chromosome translocation t(1;11)(q23;p15). Blood, 1999;2:741-7. 78. Zivin JA, Acute stroke therapy with tissue plasminogen activator (tPA) since it was approved by the U.S. Food and Drug Administration (FDA). Ann Neurol, 2009;1:6-10. 79. Inyang AL and Tobelem G, Tissue-plasminogen activator stimulates endothelial cell migration in wound assays. Biochem Biophys Res Commun, 1990;3:1326-32. 80. Ling C, Zou T, Hsiao Y, et al., Disruption of tissue plasminogen activator gene reduces macrophage migration. Biochem Biophys Res Commun, 2006;3:906-12. 81. Sugimura M, Kobayashi H, Kanayama N, et al., Clinical significance of urokinase-type plasminogen activator (uPA) in invasive cervical cancer of the uterus. Gynecol Oncol, 1992;3:330-6. 82. Shariat SF, Roehrborn CG, McConnell JD, et al., Association of the circulating levels of the urokinase system of plasminogen activation with the presence of prostate cancer and invasion, progression, and metastasis. J Clin Oncol, 2007;4:349-55. 83. Bauer TW, Liu W, Fan F, et al., Targeting of urokinase plasminogen activator receptor in human pancreatic carcinoma cells inhibits c-Met- and insulin-like growth factor-I receptor-mediated migration and invasion and orthotopic tumor growth in mice. Cancer Res, 2005;17:7775-81. 84. Duffy MJ, Urokinase plasminogen activator and its inhibitor, PAI-1, as prognostic markers in breast cancer: from pilot to level 1 evidence studies. Clin Chem, 2002;8:1194-7. 85. Choong PF and Nadesapillai AP, Urokinase plasminogen activator system: a multifunctional role in tumor progression and metastasis. Clin Orthop Relat Res, 2003;415 Suppl:S46-58. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/57421 | - |
dc.description.abstract | 轉化生長因子-β (TGF-β)是一種多功能的細胞激素,它可以調控細胞生長、細胞譜系決定、細胞分化、細胞凋亡、細胞移動以及細胞貼附。它也可以引發細胞上皮-間質轉型和轉移,在腫瘤進程中扮演重要角色。在本篇實驗中,我們使用人類乳腺上皮細胞株MCF10A來探討轉化生長因子-β如何去引發細胞侵犯。我們發現在MCF10A細胞株中,轉化生長因子-β可以促進paired related homeobox 2 (Prrx2)的表現,此促進是經由依賴smad和不依賴smad途徑。讓MCF10A細胞株穩定表現Prrx2會增加細胞侵犯以及惡性轉化的能力。利用微陣列基因表現分析,我們發現當穩定表現Prrx2時,tissue plasminogen activator (tPA)的表現量也會跟著上升。根據螢光素酶報告基因檢測系統,Prrx2可以激活tPA啟動子的活性。染色質免疫沉澱實驗證明Prrx2可以直接結合在tPA的啟動子上。我們的實驗顯示,Prrx2在人類乳腺細胞中可能藉由tPA來調控轉化生長因子-β引發的腫瘤侵犯。 | zh_TW |
dc.description.abstract | Transforming growth factor-β (TGF-β) is a multifunctional cytokine which controls cell proliferation, lineage determination, differentiation, apoptosis, motility, and adhesion. It also can induce epithelial-mesenchymal transition and metastasis, and is important in cancer progression. In this study, we used human mammary epithelial cell line MCF10A to investigate how TGF-β induces cell invasion. We found that TGF-β induces expression of paired related homeobox 2 (Prrx2) through both Smad-dependent and Smad-independent pathways in MCF10A. MCF10A stably transfected with Prrx2-expressing plasmid (MCF10A-Prrx2) had enhanced invasion and malignant transformation as compared to vector control. Using microarray analysis, we found that expression of tissue plasminogen activator (tPA) was induced in MCF10A-Prrx2, compared to vector control. Cotransfection of Prrx2-expressing plasmid and tPA reporter in Human Embryonic Kidney (HEK) 293T showed that tPA promoter activity was increased as evidenced by luciferase reporter assay. Chromatin immunoprecipitation assay showed that tPA is a direct target for Prrx2. Our study suggests that Prrx2 mediates TGF-β-induced cell invasion, possibly involving tPA, in human mammary epithelial cells. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T06:45:26Z (GMT). No. of bitstreams: 1 ntu-103-R01444006-1.pdf: 3236637 bytes, checksum: 590144f82c6634ce232229c7609debf6 (MD5) Previous issue date: 2014 | en |
dc.description.tableofcontents | 口試委員審定書……………………………………………I
謝辭……………………………………………II 中文摘要……………………………………………III Abstract……………………………………………IV Contents……………………………………………V 1. Introduction……………………………………………1 1.1 Breast cancer……………………………………………1 1.2 Epithelial-mesenchymal transition…………………………2 1.3 Transforming growth factor-β………………………………4 1.4 Structure of TGF-β………………………………………………4 1.5 TGF-β signal pathway……………………………………………5 1.6 TGF-β induces EMT………………………………………………6 1.7 TGF-β induces invasion………………………………………7 1.8 Paired related homeobox 2………………………………………7 1.9 The study aim……………………………………………8 2. Materials and Methods………………………………………9 2.1 Materials………………………………………………………9 2.2 Cell culture……………………………………………………9 2.3 Plasmids and lentiviral …………………………………9 2.4 RNA isolation……………………………………………………10 2.5 RT- PCR……………………………………………………10 2.6 Real-time PCR……………………………………………………11 2.7 Western blot……………………………………………………11 2.8 MTT assay……………………………………………………12 2.9 In vitro Boyden chamber invasion and migration assay………………………………………12 2.10 Anchorage-independent growth assay………………………………………13 2.11 Three-dimensional basement membrane culture………………………………………13 2.12 Construct for tPA promoter reporter plasmid………………………………………14 2.13 Reporter assay………………………………………………………………………14 2.14 Chromatin immunoprecipitation assay (ChIP)………………………………………15 2.15 Immunofluorescence………………………………………16 2.16 RNA interference………………………………………………………………………17 3. Results………………………………………………………………………22 3.1 TGF-β induces EMT in MCF10A………………………………………22 3.2 Validation of differential gene expression between TGF-β treated or untreated MCF10A………………………22 3.3 TGF-β induces the expression of Prrx2 in MCF10A………………………………………22 3.4 Overexpression of Prrx2 does not affect cell proliferation………………………………………23 3.5 Overexpression of Prrx2 promotes cell invasion and migration………………………………………24 3.6 Overexpression of Prrx2 promotes tumorgenesis………………………………………24 3.7 The expression of EMT markers in Prrx2-overexpressed cells………………………………………24 3.8 Validation of differential gene expression between Prrx2-overexpressed cells and control cells………25 3.9 Tissue plasminogen activator (tPA) is activated by Prrx2………………………………………25 3.10 Prrx2 directly binds to the tPA promoter………………………………………25 3.11 Knockdown tPA reduced the invasion and migration enhanced by Prrx2…………………26 4. Discussion 27 4.1 Cytokines and growth factors initiate EMT………………………………………27 4.2 TGF-β regulates gene expression………………………………………28 4.3 Prrx2 is induced by TGF-β and mediates cell invasion, migration and tumorigenesis………………………28 4.4 Prrx2 in cancer………………………………………30 4.5 tPA influences the cell invasion and migration………………………………………30 5. Figures and Tables………………………………………32 Figure 1. TGF-β induces EMT in MCF10A………………………………………32 Figure 2. Validation of the genes regulated by TGF-β identified by microarray analysis ………34 Figure 3. TGF-β induces the expression of Prrx2 in MCF10A.....................................36 Figure 4. Overexpression of Prrx2 does not affect cell proliferation.............................38 Figure 5. Overexpression of Prrx2 promotes cell invasion and migration....................40 Figure 6. Overexpression of Prrx2 promotes tumorigenesis...............42 Figure 7. The expression of EMT markers in Prrx2-overexpressed cells......................44 Figure 8. Validation of the genes regulated by Prrx2 by real-time PCR........................47 Figure 9. Tissue plasminogen activator (tPA) is activated by Prrx2..............................48 Figure 10. Prrx2 directly binds to tPA promoter........................49 Figure 11. Knockdown tPA reduced the invasion and migration enhanced by Prrx2...51 Table 1. The primers used for PCR reaction………………………………………18 Table 2. The primers used for PCR reaction………………………………………19 Table 3. The primers used for PCR reaction……………………………………….20 Table 4. The primers used for ChIP………………………………………21 Table 5. TGF-β-induced genes detected by microarray...................33 Table 6. Genes regluted by Prrx2 idetified by microarray analysis..........46 7. Reference………………………………………53 | |
dc.language.iso | en | |
dc.title | Prrx2在人類乳腺細胞中調控TGF-β引發的腫瘤侵犯 | zh_TW |
dc.title | Prrx2 mediates TGF-β-induced tumor invasion in human mammary epithelial cell | en |
dc.type | Thesis | |
dc.date.schoolyear | 102-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 連晃駿(Huang-Chun Lien) | |
dc.contributor.oralexamcommittee | 陳彥榮(Yen-Rong Chen),周涵怡(Han-Yi Elizabeth Chou) | |
dc.subject.keyword | 轉化生長因子-β,Prrx2,tPA, | zh_TW |
dc.subject.keyword | TGF-β,Prrx2,tPA, | en |
dc.relation.page | 64 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2014-07-28 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 病理學研究所 | zh_TW |
顯示於系所單位: | 病理學科所 |
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