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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 黃敏銓 | |
dc.contributor.author | Yi-Ling Wu | en |
dc.contributor.author | 吳易玲 | zh_TW |
dc.date.accessioned | 2021-06-15T05:00:01Z | - |
dc.date.available | 2014-09-09 | |
dc.date.copyright | 2010-09-09 | |
dc.date.issued | 2010 | |
dc.date.submitted | 2010-07-28 | |
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Nat Rev Mol Cell Biol 2, 127-137 (2001). 37 Tong, W. M., Ellinger, A., Sheinin, Y. & Cross, H. S. Epidermal Growth Factor Receptor Expression in Primary Cultured Human Colorectal Carcinoma Cells. Br. J. Cancer 77, 1792-1798 (1998). 38 Messa, C., Russo, F., Caruso, M. G. & Leo, A. D. EGF, TGF-α, and EGF-R in Human Colorectal Adenocarcinoma. Acta Oncol. 37, 285 - 289 (1998). 39 Spano, J.-P., Lagorce, C., Atlan, D., Milano, G., Domont, J., Benamouzig, R., Attar, A., Benichou, J., Martin, A., Morere, J.-F., Raphael, M., Penault-Llorca, F., Breau, J.-L., Fagard, R., Khayat, D., & Wind, P. Impact of EGFR Expression on Colorectal Cancer Patient Prognosis and Survival. Ann. Oncol. 16, 102-108 (2005). 40 Guo, P., Wang, Q. Y., Guo, H. B., Shen, Z. H. & Chen, H. L. N-acetylglucosaminyltransferase V Modifies the Signaling Pathway of Epidermal Growth Factor Receptor. Cell. Mol. Life Sci. 61, 1795-1804 (2004). 41 Matsumoto K., Yokote H., Arao T., Maegawa M., Tanaka K., Fujita Y., Shimizu C., Hanafusa T., Fujiwara Y., & Nishio K. N-Glycan Fucosylation of Epidermal Growth Factor Receptor Modulates Receptor Activity and Sensitivity to Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor. Cancer Sci. 99, 1611-1617 (2008). 42 Liotta, L. A. & Stetler-Stevenson, W. G. Tumor Invasion and Metastasis: an Imbalance of Positive and Negative Regulation. Cancer Research 51, 5054s-5059s (1991). 43 Guo, W. & Giancotti, F. G. Integrin Signalling during Tumour Progression. Nat. Rev. Mol. Cell Biol. 5, 816-826 (2004). 44 Schaller, M. D., Hildebrand, J. D., Shannon, J. D., Fox, J. W., Vines, R. R., & Parsons, J. T. Autophosphorylation of the focal adhesion kinase, pp125FAK, directs SH2-dependent binding of pp60src. Mol. Cell. Biol. 14, 1680-1688 (1994). 45 Guo, H.-B., Lee, I., Kamar, M., Akiyama, S. K. & Pierce, M. Aberrant N-glycosylation of β1 Integrin Causes Reduced α5β1 Integrin Clustering and Stimulates Cell Migration. Cancer Research 62, 6837-6845 (2002). 46 Zhuo, Y., Chammas, R. & Bellis, S. L. Sialylation of β1 Integrins Blocks Cell Adhesion to Galectin-3 and Protects Cells against Galectin-3-induced Apoptosis. J. Biol. Chem. 283, 22177-22185 (2008). 47 Cummings, R. D., Soderquist, A. M. & Carpenter, G. The Oligosaccharide Moieties of the Epidermal Growth Factor Receptor in A-431 cells. Presence of Complex-type N-linked Chains that Contain Terminal N-acetylgalactosamine Residues. J. Biol. Chem. 260, 11944-11952 (1985). 48 Gu, J., Isaji, T., Sato, Y., Kariya, Y. & Fukuda, T. Importance of N-glycosylation on α5β1 Integrin for its Biological Functions. Biol. Pharm. Bull. 32, 780-785 (2009). | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/46251 | - |
dc.description.abstract | β4半乳糖轉移酵素(B4GALTs)能夠將雙磷酸尿苷-半乳糖(UDP-galactose)經由β-1,4-糖苷鍵結接到氨基乙醯葡萄糖苷(N-acetylglucosamine)之上而生成氨基乙醯乳糖苷(N-acetyllactosamine)。重複的氨基乙醯乳糖苷單元可組成的聚氨基乙醯乳糖苷(poly-N-acetyllactosamine)。在本研究中,我們闡明了β4半乳糖轉移酵素三號 (B4GALT3)在HCT116大腸癌細胞之功能性角色。過度表達B4GALT3促進了細胞在體外的增殖以及老鼠體內腫瘤的生長;同時,將細胞培養在懸浮環境時,過度表達B4GALT3也促進了細胞群落的形成。另外,過度表達 B4GALT3也降低了細胞遷移和侵襲能力。有趣的是,B4GALT3會增加表皮生長因子受體(EGFR)以及β1整合素(β1 integrin)上的poly-N-acetyllactosamine結構。在過度表達 B4GALT3的細胞中,我們發現了EGFR的845酪胺酸位置有較多的磷酸化現象(pY845),這可能是促使細胞生長增加的原因。此外,B4GALT3會降低FAK和paxillin的磷酸化,這表示了導致細胞遷移能力下降的部分原因可能是藉由integrin信號傳導通路。經由免疫組織化學染色我們發現了在64.6% (7/11)大腸直腸癌病人中,B4GALT3是表現量上升的;然而,經由西方點墨法及定量聚合酵素鏈鎖反應,卻發現分別只有40% (4/10)和44.6% (37/83)的病人有B4GALT3表現量上升的現象。這種不一致性可能是由於一些在大腸黏膜的基質細胞也表現有B4GALT3所造成。 | zh_TW |
dc.description.abstract | β-1,4-galactosyltransferases (B4GALTs) catalyze the transfer of UDP-galactose to an N-acetylglucosamine via β-1,4-linkage and are responsible for the biosynthesis of N-acetyllactosamine. Poly-N-acetyllactosamines are composed of repeating units of N-acetyllactosamine. In the present study, we elucidate the function of B4GALT3 in HCT116 colon cancer cells. Overexpression of B4GALT3 promotes cell proliferation in vitro and tumor growth in vivo, as well as colony formation. Overexpression of B4GALT3 also reduces cell migration and invasion. Interestingly, B4GALT3 increases poly-N-acetyllactosamine structures on both EGFR and β1 integrin. Up-regulation of phosphorylated EGFR (pY845) is found in cells overexpressing B4GALT3 and this may lead to an increase in cell growth. Moreover, B4GALT3 decreases phosphorylation of focal adhesion kinase (FAK) and paxillin, indicating that reduced cell migration may be partly mediated by integrin signaling pathways. Immunohistochemistry shows that B4GALT3 is up-regulated in 64.6% (7/11) of colorectal tumors compared with their normal counterparts, whereas only 40% (4/10) and 44.6% (37/83) detected by Western blotting and QPCR, respectively. This inconsistency may result from the positive staining of some stromal cells in colorectal mucosae. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T05:00:01Z (GMT). No. of bitstreams: 1 ntu-99-R97446006-1.pdf: 1842534 bytes, checksum: 76ff353d3d95d5e01682247668101f2c (MD5) Previous issue date: 2010 | en |
dc.description.tableofcontents | 口試委員會審定書 3
中文摘要 4 Abstract 5 Chapter 1 Introduction 6 Chapter 2 Materials and Methods 11 2.1 Cell Culture and Transfection 11 2.2 Colorectal Tissue Samples 11 2.3 RNA Isolation, Reverse Transcription, and QPCR 12 2.4 MTT Assay 13 2.5 Tumor Growth in SCID Mice 13 2.6 Anchorage-independent Growth in Soft Agar (Colony Formation Assay) 14 2.7 Migration Assay 14 2.8 Invasion Assay 15 2.9 Western Blotting 15 2.10 LEL Pull-down 16 2.11 Deglycosylation 17 2.12 Immunohistochemistry 17 Chapter 3 Results 19 3.1 Expression Levels of B4GALTs in Human Normal Colon Tissues 19 3.2 Expression of B4GALT3 in Colon Cancer Cells 19 3.3 Overexpression of B4GALT3 in HCT116 Cells 20 3.4 B4GALT3 Promotes Cell Growth in HCT116 Cells 21 3.5 B4GALT3 Enhances Tumor Growth in SCID Mice 21 3.6 B4GALT3 Increases Colony-forming Abilities in HCT116 Cells 22 3.7 B4GALT3 Suppresses Migratory and Invasive Abilities in HCT116 Cells 22 3.8 B4GALT3 Suppresses Downstream Signaling of Integrins 23 3.9 B4GALT3 Alters Glycosylation States of β1 Integrin 23 3.10 The Decoration of Poly-N-acetyllactosamine is Mainly on O-glycans of β1 Integrin 24 3.11 B4GALT3 Alters Glycosylation States of EGFR and Enhances Its Phosphorylation at Y845 25 3.12 The Position of Poly-N-acetyllactosaminyl Decoration on EGFR is Unknown 25 3.13 Expression of B4GALT3 mRNA in CRC Patients 26 3.14 Expression of B4GALT3 in CRC Patients by Western Blotting 26 3.15 B4GALT3 is Up-regulated in Colorectal Tumor Tissues 27 Chapter 4 Discussion 28 Reference 32 Figures and Tables 40 Table 1. Primers used for QPCR analysis 41 Table 2. H Scores of immunohistochemistry 42 Figure 1 43 Figure 2 44 Figure 3 45 Figure 4 46 Figure 5 47 Figure 6 48 Figure 7 49 Figure 8 50 Figure 9 51 Figure 10 52 Figure 11 53 Figure 12 54 Figure 13. 55 Appendix 56 Abbreviations 57 | |
dc.language.iso | en | |
dc.title | B4GALT3在大腸癌細胞中的功能性角色 | zh_TW |
dc.title | Functional Roles of B4GALT3 in Colon Cancer Cells | en |
dc.type | Thesis | |
dc.date.schoolyear | 98-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 李財坤,李明學 | |
dc.subject.keyword | β4半乳糖轉移酵素,大腸直腸癌,表皮生長因子受體,β1 整合素, | zh_TW |
dc.subject.keyword | β1,4-galactosyltransferase,colorectal cancer,EFGR,β1 integrin, | en |
dc.relation.page | 58 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2010-07-29 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 解剖學暨生物細胞學研究所 | zh_TW |
顯示於系所單位: | 解剖學暨細胞生物學科所 |
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