請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/25099
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 王萬波 | |
dc.contributor.author | Shu-Fan Chou | en |
dc.contributor.author | 周書帆 | zh_TW |
dc.date.accessioned | 2021-06-08T06:02:16Z | - |
dc.date.copyright | 2007-08-08 | |
dc.date.issued | 2007 | |
dc.date.submitted | 2007-07-26 | |
dc.identifier.citation | Abid, M.R., Yi, X., Yano, K., Shih, S.C. and Aird, W.C. (2006) Vascular endocan is preferentially expressed in tumor endothelium. Microvasc Res, 72, 136-145.
Aitkenhead, M., Wang, S.J., Nakatsu, M.N., Mestas, J., Heard, C. and Hughes, C.C. (2002) Identification of endothelial cell genes expressed in an in vitro model of angiogenesis: induction of ESM-1, (beta)ig-h3, and NrCAM. Microvasc Res, 63, 159-171. Allen, S.J., Crown, S.E. and Handel, T.M. (2007) Chemokine: receptor structure, interactions, and antagonism. Annu Rev Immunol, 25, 787-820. Bechard, D., Gentina, T., Delehedde, M., Scherpereel, A., Lyon, M., Aumercier, M., Vazeux, R., Richet, C., Degand, P., Jude, B., Janin, A., Fernig, D.G., Tonnel, A.B. and Lassalle, P. (2001a) Endocan is a novel chondroitin sulfate/dermatan sulfate proteoglycan that promotes hepatocyte growth factor/scatter factor mitogenic activity. J Biol Chem, 276, 48341-48349. Bechard, D., Meignin, V., Scherpereel, A., Oudin, S., Kervoaze, G., Bertheau, P., Janin, A., Tonnel, A. and Lassalle, P. (2000) Characterization of the secreted form of endothelial-cell-specific molecule 1 by specific monoclonal antibodies. J Vasc Res, 37, 417-425. Bechard, D., Scherpereel, A., Hammad, H., Gentina, T., Tsicopoulos, A., Aumercier, M., Pestel, J., Dessaint, J.P., Tonnel, A.B. and Lassalle, P. (2001b) Human endothelial-cell specific molecule-1 binds directly to the integrin CD11a/CD18 (LFA-1) and blocks binding to intercellular adhesion molecule-1. J Immunol, 167, 3099-3106. Bikfalvi, A., Klein, S., Pintucci, G. and Rifkin, D.B. (1997) Biological roles of fibroblast growth factor-2. Endocr Rev, 18, 26-45. Borczuk, A.C., Shah, L., Pearson, G.D., Walter, K.L., Wang, L., Austin, J.H., Friedman, R.A. and Powell, C.A. (2004) Molecular signatures in biopsy specimens of lung cancer. Am J Respir Crit Care Med, 170, 167-174. Chen, C.Y. and Shyu, A.B. (1995) AU-rich elements: characterization and importance in mRNA degradation. Trends Biochem Sci, 20, 465-470. Cong, R., Jiang, X., Wilson, C.M., Hunter, M.P., Vasavada, H. and Bogue, C.W. (2006) Hhex is a direct repressor of endothelial cell-specific molecule 1 (ESM-1). Biochem Biophys Res Commun, 346, 535-545. Delehedde, M., Allain, F.,Payne, S.J.,Borgo, R.,Vanpouille, C.,Fernig, D.G., and Deudon, E. (2002) Proteoglycans in Inflammation. Current Medicinal Chemistry - Anti-Inflammatory & Anti-Allergy Agents, 1, 89-102. Delehedde, M., Lyon, M., Vidyasagar, R., McDonnell, T.J. and Fernig, D.G. (2002) Hepatocyte growth factor/scatter factor binds to small heparin-derived oligosaccharides and stimulates the proliferation of human HaCaT keratinocytes. J Biol Chem, 277, 12456-12462. Duan, H., Cheng, L., Sun, X., Wu, Y., Hu, L., Wang, J., Zhao, H. and Lu, G. (2006) LFA-1 and VLA-4 involved in human high proliferative potential-endothelial progenitor cells homing to ischemic tissue. Thromb Haemost, 96, 807-815. Ferrara, N., Gerber, H.P. and LeCouter, J. (2003) The biology of VEGF and its receptors. Nat Med, 9, 669-676. Folkman, J. (1971) Tumor angiogenesis: therapeutic implications. N Engl J Med, 285, 1182-1186. Folkman, J. (1995) Angiogenesis in cancer, vascular, rheumatoid and other disease. Nat Med, 1, 27-31. Frevel, M.A., Bakheet, T., Silva, A.M., Hissong, J.G., Khabar, K.S. and Williams, B.R. (2003) p38 Mitogen-activated protein kinase-dependent and -independent signaling of mRNA stability of AU-rich element-containing transcripts. Mol Cell Biol, 23, 425-436. Gerritsen, M.E., Tomlinson, J.E., Zlot, C., Ziman, M. and Hwang, S. (2003) Using gene expression profiling to identify the molecular basis of the synergistic actions of hepatocyte growth factor and vascular endothelial growth factor in human endothelial cells. Br J Pharmacol, 140, 595-610. Gory, S., Dalmon, J., Prandini, M.H., Kortulewski, T., de Launoit, Y. and Huber, P. (1998) Requirement of a GT box (Sp1 site) and two Ets binding sites for vascular endothelial cadherin gene transcription. J Biol Chem, 273, 6750-6755. Grant, D.S., Yenisey, C., Rose, R.W., Tootell, M., Santra, M. and Iozzo, R.V. (2002) Decorin suppresses tumor cell-mediated angiogenesis. Oncogene, 21, 4765-4777. Grigoriu, B.D., Depontieu, F., Scherpereel, A., Gourcerol, D., Devos, P., Ouatas, T., Lafitte, J.J., Copin, M.C.,Tonnel, A.B. and Lassalle, P. (2006) Endocan expression and relationship with survival in human non-small cell lung cancer. Clin Cancer Res, 12, 4575-4582. Gupta, K. and Zhang, J. (2005) Angiogenesis: a curse or cure? Postgrad Med J, 81, 236-242. Hendrix, M.J., Seftor, E.A., Hess, A.R. and Seftor, R.E. (2003) Molecular plasticity of human melanoma cells. Oncogene, 22, 3070-3075. Iozzo, R.V. (1998) Matrix proteoglycans: from molecular design to cellular function. Annu Rev Biochem, 67, 609-652. Iozzo, R.V. and Cohen, I. (1993) Altered proteoglycan gene expression and the tumor stroma. Experientia, 49, 447-455. Iozzo, R.V. and San Antonio, J.D. (2001) Heparan sulfate proteoglycans: heavy hitters in the angiogenesis arena. J Clin Invest, 108, 349-355. Ish-Horowicz, D. and Burke, J.F. (1981) Rapid and efficient cosmid cloning. Nucleic Acids Res, 9, 2989-2998. Kerbel, R. and Folkman, J. (2002) Clinical translation of angiogenesis inhibitors. Nat Rev Cancer, 2, 727-739. Kerbel, R.S. (2000) Tumor angiogenesis: past, present and the near future. Carcinogenesis, 21, 505-515. Kirkpatrick, C.A. and Selleck, S.B. (2007) Heparan sulfate proteoglycans at a glance. J Cell Sci, 120, 1829-1832. Kjellen, L. and Lindahl, U. (1991) Proteoglycans: structures and interactions. Annu Rev Biochem, 60, 443-475. Kramer, K.L. and Yost, H.J. (2003) Heparan sulfate core proteins in cell-cell signaling. Annu Rev Genet, 37, 461-484. Lassalle, P., Molet, S., Janin, A., Heyden, J.V., Tavernier, J., Fiers, W., Devos, R. and Tonnel, A.B. (1996) ESM-1 is a novel human endothelial cell-specific molecule expressed in lung and regulated by cytokines. J Biol Chem, 271, 20458-20464. Lenburg, M.E., Liou, L.S., Gerry, N.P., Frampton, G.M., Cohen, H.T. and Christman, M.F. (2003) Previously unidentified changes in renal cell carcinoma gene expression identified by parametric analysis of microarray data. BMC Cancer, 3, 31. Li, A., Dubey, S., Varney, M.L., Dave, B.J. and Singh, R.K. (2003) IL-8 directly enhanced endothelial cell survival, proliferation, and matrix metalloproteinases production and regulated angiogenesis. J Immunol, 170, 3369-3376. Liekens, S., De Clercq, E. and Neyts, J. (2001) Angiogenesis: regulators and clinical applications. Biochem Pharmacol, 61, 253-270. Lyon, M., Deakin, J.A. and Gallagher, J.T. (2002) The mode of action of heparan and dermatan sulfates in the regulation of hepatocyte growth factor/scatter factor. J Biol Chem, 277, 1040-1046. Lyon, M., Deakin, J.A., Rahmoune, H., Fernig, D.G., Nakamura, T. and Gallagher, J.T. (1998) Hepatocyte growth factor/scatter factor binds with high affinity to dermatan sulfate. J Biol Chem, 273, 271-278. Oikawa, T. and Yamada, T. (2003) Molecular biology of the Ets family of transcription factors. Gene, 303, 11-34. Pandya, N.M., Dhalla, N.S. and Santani, D.D. (2006) Angiogenesis--a new target for future therapy. Vascul Pharmacol, 44, 265-274. Penc, S.F., Pomahac, B., Winkler, T., Dorschner, R.A., Eriksson, E., Herndon, M. and Gallo, R.L. (1998) Dermatan sulfate released after injury is a potent promoter of fibroblast growth factor-2 function. J Biol Chem, 273, 28116-28121. Ramsauer, M. and D'Amore, P.A. (2002) Getting Tie(2)d up in angiogenesis. J Clin Invest, 110, 1615-1617. Reed, C.C., Waterhouse, A., Kirby, S., Kay, P., Owens, R.T., McQuillan, D.J. and Iozzo, R.V. (2005) Decorin prevents metastatic spreading of breast cancer. Oncogene, 24, 1104-1110. Rennel, E., Mellberg, S., Dimberg, A., Petersson, L., Botling, J., Ameur, A., Westholm, J.O., Komorowski, J., Lassalle, P., Cross, M.J. and Gerwins, P. (2007) Endocan is a VEGF-A and PI3K regulated gene with increased expression in human renal cancer. Exp Cell Res, 313, 1285-1294. Rhee, J. and Hoff, P.M. (2005) Angiogenesis inhibitors in the treatment of cancer. Expert Opin Pharmacother, 6, 1701-1711. Rhim, J.S., Jay, G., Arnstein, P., Price, F.M., Sanford, K.K. and Aaronson, S.A. (1985) Neoplastic transformation of human epidermal keratinocytes by AD12-SV40 and Kirsten sarcoma viruses. Science, 227, 1250-1252. Scherpereel, A., Depontieu, F., Grigoriu, B., Cavestri, B., Tsicopoulos, A., Gentina, T., Jourdain, M., Pugin, J., Tonnel, A.B. and Lassalle, P. (2006) Endocan, a new endothelial marker in human sepsis. Crit Care Med, 34, 532-537. Scherpereel, A., Gentina, T., Grigoriu, B., Senechal, S., Janin, A., Tsicopoulos, A., Plenat, F., Bechard, D., Tonnel, A.B. and Lassalle, P. (2003) Overexpression of endocan induces tumor formation. Cancer Res, 63, 6084-6089. Seidel, C., Borset, M., Hjertner, O., Cao, D., Abildgaard, N., Hjorth-Hansen, H., Sanderson, R.D., Waage, A. and Sundan, A. (2000) High levels of soluble syndecan-1 in myeloma-derived bone marrow: modulation of hepatocyte growth factor activity. Blood, 96, 3139-3146. Takeshita, H., Yoshizaki, T., Miller, W.E., Sato, H., Furukawa, M., Pagano, J.S. and Raab-Traub, N. (1999) Matrix metalloproteinase 9 expression is induced by Epstein-Barr virus latent membrane protein 1 C-terminal activation regions 1 and 2. J Virol, 73, 5548-5555. Timar, J., Lapis, K., Dudas, J., Sebestyen, A., Kopper, L. and Kovalszky, I. (2002) Proteoglycans and tumor progression: Janus-faced molecules with contradictory functions in cancer. Semin Cancer Biol, 12, 173-186. Trowbridge, J.M. and Gallo, R.L. (2002) Dermatan sulfate: new functions from an old glycosaminoglycan. Glycobiology, 12, 117R-125R. Tsai, J.C., Zhang, J., Minami, T., Voland, C., Zhao, S., Yi, X., Lassalle, P., Oettgen, P. and Aird, W.C. (2002) Cloning and characterization of the human lung endothelial-cell-specific molecule-1 promoter. J Vasc Res, 39, 148-159. Tschernatsch, M.M., Mlecnik, B., Trajanoski, Z., Zechner, R. and Zimmermann, R. (2006) LPL-mediated lipolysis of VLDL induces an upregulation of AU-rich mRNAs and an activation of HuR in endothelial cells. Atherosclerosis, 189, 310-317. van 't Veer, L.J., Dai, H., van de Vijver, M.J., He, Y.D., Hart, A.A., Mao, M., Peterse, H.L., van der Kooy, K., Marton, M.J., Witteveen, A.T., Schreiber, G.J., Kerkhoven, R.M., Roberts, C., Linsley, P.S., Bernards, R. and Friend, S.H. (2002) Gene expression profiling predicts clinical outcome of breast cancer. Nature, 415, 530-536. Wakiya, K., Begue, A., Stehelin, D. and Shibuya, M. (1996) A cAMP response element and an Ets motif are involved in the transcriptional regulation of flt-1 tyrosine kinase (vascular endothelial growth factor receptor 1) gene. J Biol Chem, 271, 30823-30828. Wellner, M., Herse, F., Janke, J., Gorzelniak, K., Engeli, S., Bechart, D., Lasalle, P., Luft, F.C. and Sharma, A.M. (2003) Endothelial cell specific molecule-1--a newly identified protein in adipocytes. Horm Metab Res, 35, 217-221. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/25099 | - |
dc.description.abstract | Endocan(又名內皮細胞特異表現分子,Esm-1)為一分子量約50 kDa的可溶性蛋白多醣,由核心蛋白與一條condroitin/ dermatan sulfate(CS/DS)醣基鏈組成,主要表現於人體肺、腎組織及內皮細胞中。研究顯示,內皮細胞的endocan會受到週邊環境因子的調控;如發炎時大量表現的細胞激素TNF-α,及促進細胞增生、血管新生的生長因子VEGF-A等。前人之in vitro實驗證實,endocan可與肝細胞生長因子HGF/SF共同作用,藉其醣基鏈加強HGF/SF促進細胞增生的能力。文獻指出,於老鼠體內移植穩定表現endocan之HEK293細胞後,易觀察到腫瘤生成現象;其中核心蛋白胺基酸F115-116序列及醣基鏈對於endocan促進腫瘤生長的功能而言十分重要。臨床研究也發現,在許多癌症及敗血休克病人的血液中皆可偵測到高表現量的endocan;其表現量與癌症病人之癒後恢復不良及血管新生現象息息相關,故推測endocan可做為腫瘤嚴重程度的指標。以上研究顯示,endocan可能影響腫瘤生長及伴隨而來的血管新生現象。
為了進一步確定endocan與細胞癌化的關聯性,本研究首先建立穩定表現endocan的RHEK-1細胞株,利用相關in vitro 實驗探討其生長特性、細胞爬行能力及致癌潛能。結果顯示,穩定表現endocan的RHEK-1細胞株不論是生長速率、細胞爬行能力或細胞聚落形成能力,相較母株細胞或載體表現細胞株皆無明顯差異,此一結果顯示endocan似乎無法直接影響細胞癌化路徑。 然而在血管新生的研究上,以含少量血清之endocan條件化上清液培養人類微血管內皮細胞株HMEC-1,偵測內皮細胞增生情形;結果顯示,endocan與少量血清共同作用時可促進內皮細胞生長。此外,含大量endocan之無血清條件化上清液可有效吸引HMEC-1與人類臍帶靜脈內皮細胞HUVEC轉移;於條件化上清液加入endocan抗體後即降低轉移之HMEC-1數目,證明endocan為促進內皮細胞轉移之重要因子。利用endocan突變細胞株進行實驗,結果顯示endocan組成之醣基鏈與核心蛋白胺基酸F115-116序列為其吸引內皮細胞轉移之重要區域。後續實驗發現endocan亦可促進內皮細胞侵犯能力。綜合上述實驗結果,推測endocan參與在血管新生之特定步驟,然而其與腫瘤癌化及血管新生的關聯性仍需進一步於相關實驗中証實。 | zh_TW |
dc.description.abstract | Endocan (previously called endothelial cell-specific molecule 1, Esm-1) is a 50 kDa soluble proteoglycan that is constituted of a core protein and a single condroitin/dermatan sulfate (CS/DS)chain linked to the serine residue. Endocan is predominantly expressed and secreted by vascular endothelium in response to pro-inflammatory cytokines and pro-angiogenic factors such as TNF-α and VEGF-A. Endocan can bind to the hepatocyte growth factor/scatter factor (HGF/SF) through its glycan domain, thereby enhance HGF/SF mitogenic ability. In mouse model of human tumor xenograft, endocan expression can lead to an increased tumor incidence and this activity of endocan is dependent on its glycan chain moiety and F115-116 residues. Clinical studies indicated that levels of circulating endocan were elevated in several types of cancers and inflammatory disorders, and its expression was correlated with poor prognosis and severity of angiogenic status in cancer patients. In this regard, endocan can be used as a marker of tumor aggressiveness. Previous studies suggest that endocan may be involved in controlling tumor development and play a potential role in angiogenesis accompanied by tumor progression.
Since the exact role of endocan in tumorigenesis is still vague, in the study, we first established endocan-overexpressing RHEK-1 cells lines and studied their growth property, cell migration ability, and tumorigenic potential. Our data indicated that neither growth kinetics nor cell migration ability or transforming ability of these endocan-expressing cell lines were altered when compared to the vector-transfected and parental cell lines. Therefore endocan may not directly involve in the cellular transformation process. We then tested whether endocan played roles in angiogenesis. Low-serum-containing conditioned medium from endocan-expressing cells or vector-transfected cells were added to the human microvascular endothelial cells (HMEC-1)and the proliferation of HMEC-1 was measured. Our data indicated that endocan could stimulate the growth of HMEC-1 cells in the presence of low serum. Serum-free conditioned medium from endocan-expressing cells were found to be able to stimulate the migration of HMEC-1 and human umbilical vein endothelial cells (HUVEC) in the transwell migration assays. This ability of endocan-containing conditioned medium was indeed attributed to endocan because anti-endocan antibody could block the ability of conditioned medium to promote HMEC-1 migration. Mutant studies indicated that endocan core protein region surrounding F115-116 residues and its GAG chain moiety were important for endocan’s migration-stimulating function. In addition to stimulating the migration of endothelial cells, endocan was also found to be able to stimulate the invasion ability of HMEC-1. Together, these data indicate that endocan does play a role in angiogenesis. Its effect on tumor progression and angiogenesis requires further investigation. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T06:02:16Z (GMT). No. of bitstreams: 1 ntu-96-R94445106-1.pdf: 1566923 bytes, checksum: 45deb2d85e6ce787f31295d94aac7d5a (MD5) Previous issue date: 2007 | en |
dc.description.tableofcontents | 中文摘要....................................................i
Abstract................................................iii 目錄......................................................v 圖目錄.................................................viii 序言......................................................1 ㄧ、蛋白多醣(Proteoglycan)..............................1 二、內皮細胞特異表現分子(Endocan).......................1 三、Endocan之基因調控.....................................2 四、Endocan相關功能之報導.................................3 五、Endocan及相關疾病研究.................................5 六、血管新生(Angiogenesis)..............................5 研究目的..................................................7 材料與方法................................................8 ◆實驗材料................................................8 ㄧ、化學藥品及試劑......................................8 二、套組試劑...........................................10 三、抗體...............................................11 四、其他...............................................11 五、細胞株(Cell line)................................11 六、質體(Plasmid)....................................12 ◆實驗方法...............................................13 一、定點突變質體之建構(Site-directed mutagenesis)....13 二、細菌轉形(Transformation).........................14 三、小量質體的製備(Mini-preparation).................14 四、大量質體的製備(Maxi-preparation).................15 五、質體轉染(Transfection)...........................16 六、細胞核糖核酸萃取(RNA extraction).................17 七、反轉錄反應(Reverse transcription)................18 八、反轉錄聚合酶連鎖反應(RT-PCR).....................18 九、細胞全蛋白質與條件化培養液蛋白質之收取.............19 十、蛋白質定量.........................................20 十一、西方墨點法(Western blot).......................20 十二、酵素連結免疫吸附分析(Enzyme-linked immunosorbent assay, ELISA)...................................21 十三、MTT assay........................................22 十四、細胞傷口癒合實驗(Wound healing assay)..........22 十五、明膠蛋白酵素電泳法(Gelatin zymography).........23 十六、柔軟瓊脂中細胞聚落形成分析(Soft agar colony formation assay)................................23 十七、細胞轉移及侵犯實驗(Migration and invasion assay)...........................................24 十八、2D管柱形成實驗(Tube formation assay)...........24 實驗結果.................................................26 一、建構endocan穩定表現細胞株(stable clone)及表現 endocan的混合細胞株(pooled clone)................26 二、Endocan不影響RHEK-1細胞株生長速率..................27 三、Endocan不影響RHEK-1細胞間質金屬結合蛋白酶MMP-2、 MMP-9表現量........................................27 四、Endocan不影響RHEK-1細胞爬行能力....................28 五、Endocan不影響RHEK-1細胞轉形能力....................28 六、含血清之endocan條件化培養液可促進內皮細胞HMEC-1 生長...............................................28 七、含有endocan之條件化培養液可有效吸引內皮細胞HMEC-1與 HUVEC轉移..........................................29 八、經endocan抗體處理之endocan條件化培養液無法有效吸引內 皮細胞HMEC-1轉移...................................30 九、細菌表現之endocan核心蛋白無法吸引內皮細胞HMEC-1轉 移.................................................30 十、核心蛋白F115-116胺基酸與GAG鏈為endocan吸引內皮細胞轉 移之重要區域.......................................31 十一、Endocan可促進內皮細胞HMEC-1之侵犯能力............31 十二、Endocan不影響內皮細胞形成2D管柱型態..............32 討論.....................................................33 附圖.....................................................38 參考文獻.................................................50 | |
dc.language.iso | zh-TW | |
dc.title | Endocan促進內皮細胞轉移與侵犯能力之研究 | zh_TW |
dc.title | Endocan Promotes the Migration and Invasion Ability of Endothelial Cells | en |
dc.type | Thesis | |
dc.date.schoolyear | 95-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 陳振陽,鄧述諄 | |
dc.subject.keyword | 蛋白多醣,內皮細胞,轉移,血管新生, | zh_TW |
dc.subject.keyword | proteoglycan,endothelial cell,migration,angiogenesis, | en |
dc.relation.page | 54 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2007-07-27 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 微生物學研究所 | zh_TW |
顯示於系所單位: | 微生物學科所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-96-1.pdf 目前未授權公開取用 | 1.53 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。