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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 | 黃敏銓(Min-Chuan Huang) | |
dc.contributor.author | Chih-Wei Chen | en |
dc.contributor.author | 陳知瑋 | zh_TW |
dc.date.accessioned | 2021-06-13T06:45:54Z | - |
dc.date.available | 2013-10-07 | |
dc.date.copyright | 2011-10-07 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2011-07-25 | |
dc.identifier.citation | Birchmeier, C., Birchmeier, W., Gherardi, E. and Vande Woude, G. F. (2003) Met, metastasis, motility and more. Nat Rev Mol Cell Biol, 4, 915-925.
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Itoh, Y., Kamata-Sakurai, M., Denda-Nagai, K., Nagai, S., Tsuiji, M., Ishii-Schrade, K., Okada, K., Goto, A., Fukayama, M. and Irimura, T. (2008) Identification and expression of human epiglycanin/MUC21: a novel transmembrane mucin. Glycobiology, 18, 74-83. Karumanchi, S. A., Maynard, S. E., Stillman, I. E., Epstein, F. H. and Sukhatme, V. P. (2005) Preeclampsia: a renal perspective. Kidney Int, 67, 2101-2113. Kinoshita, M., Nakamura, T., Ihara, M., Haraguchi, T., Hiraoka, Y., Tashiro, K. and Noda, M. (2001) Identification of human endomucin-1 and -2 as membrane-bound O-sialoglycoproteins with anti-adhesive activity. FEBS Lett, 499, 121-126. McGuckin, M. A., Linden, S. K., Sutton, P. and Florin, T. H. (2011) Mucin dynamics and enteric pathogens. Nat Rev Microbiol, 9, 265-278. Ohlsson, R. (1989) Growth factors, protooncogenes and human placental development. Cell Differ Dev, 28, 1-15. Pallesen, L. T., Berglund, L., Rasmussen, L. K., Petersen, T. E. and Rasmussen, J. T. (2002) Isolation and characterization of MUC15, a novel cell membrane-associated mucin. Eur J Biochem, 269, 2755-2763. Pijnenborg, R., Bland, J. M., Robertson, W. B. and Brosens, I. (1983) Uteroplacental arterial changes related to interstitial trophoblast migration in early human pregnancy. Placenta, 4, 397-413.Prakobphol, A., Genbacev, O., Gormley, M., Kapidzic, M. and Fisher, S. J. (2006) A role for the L-selectin adhesion system in mediating cytotrophoblast emigration from the placenta. Dev Biol, 298, 107-117. Pijnenborg, R., Vercruysse, L. and Hanssens, M. (2008) Fetal-maternal conflict, trophoblast invasion, preeclampsia, and the red queen. Hypertens Pregnancy, 27, 183-196. Pochampalli, M. R., el Bejjani, R. M. and Schroeder, J. A. (2007) MUC1 is a novel regulator of ErbB1 receptor trafficking. Oncogene, 26, 1693-1701. Qiao, H., Saulnier, R., Patryzkat, A., Rahimi, N., Raptis, L., Rossiter, J., Tremblay, E. and Elliott, B. 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K., Zarnegar, R., Oliver, L. and Michalopoulos, G. K. (1991) Hepatocyte growth factor in human placenta and trophoblastic disease. Am J Pathol, 138, 1035-1043. Staun-Ram, E., Goldman, S., Gabarin, D. and Shalev, E. (2004) Expression and importance of matrix metalloproteinase 2 and 9 (MMP-2 and -9) in human trophoblast invasion. Reprod Biol Endocrinol, 2, 59. Uehara, Y., Minowa, O., Mori, C., Shiota, K., Kuno, J., Noda, T. and Kitamura, N. (1995) Placental defect and embryonic lethality in mice lacking hepatocyte growth factor/scatter factor. Nature, 373, 702-705. Weidner, K. M., Behrens, J., Vandekerckhove, J. and Birchmeier, W. (1990) Scatter factor: molecular characteristics and effect on the invasiveness of epithelial cells. J Cell Biol, 111, 2097-2108. Zhou, Y., Fisher, S. J., Janatpour, M., Genbacev, O., Dejana, E., Wheelock, M. and Dekker, G. A. and Sibai, B. M. (1998) Etiology and pathogenesis of preeclampsia: current concepts. Am J Obstet Gynecol, 179, 1359-1375. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/35262 | - |
dc.description.abstract | 在人類胎盤的發育過程中滋養層母細胞(trophoblast)的侵襲作用(invasion)影響了後續胎兒是否能正常生長。在前人研究中指出黏液蛋白(Mucin)表現在許多的上皮細胞(epithelial cells),其中也包含了滋養層母細胞,而其表現對細胞的生理有重要的影響。胎盤中部分滋養層母細胞會分泌肝細胞生長因子(Hepatocyte growth factor),此物質在胎盤中大量存在同時也被報導對於胎盤的發育有重大的影響。
在我們先前的研究中發現數種黏液蛋白的確表現在胎盤中且對於胎盤的發育有重要的影響。MUC20屬於黏液蛋白家族的成員,由人類腎臟基因庫中被鑑定出來,發現其在人類腎臟有大量的表現外,在胎盤、大腸、肺臟、前列腺和肝臟也有一定的表現量。因此,探討MUC20在胎盤中的表現以及其對滋養層母細胞生理功能的影響,可以讓我們對於黏液蛋白在胎盤發育過程中所扮演的角色有更多的瞭解。 在本研究中由西方墨點法分析發現MUC20在第一孕程(1-3個月)表現量最高,第二孕程(4-6個月)時表現量最少,第三孕程(7-9個月)表現量介於第一和第二孕程之間。再使用IHC染色得到MUC20在第一孕程有大量的表現,其中主要表現在細胞索(cell column)和絨毛的胞融型滋養層母細胞(syncytiotrophoblast),但細胞型滋養層母細胞(cytotrophoblast)和絨毛外滋養層母細胞(extravillous trophoblast)則沒有表現;此外,在第三孕程的絨毛外滋養層母細胞則出現了MUC20的表現,而此時期的胞融型滋養層母細胞也有微量的表現。接著觀察MUC20表現在類滋養層母細胞株JAR和3A-Sub-E時對細胞爬行(migration)和侵襲能力的影響。發現在以血清吸引的條件下,當MUC20表現量增加時會抑制細胞的爬行和侵襲能力;在以肝細胞生長因子吸引的條件下也會抑制細胞的侵襲能力,但不能造成細胞的爬行。最後透過肝細胞生長因子刺激細胞研究細胞內的訊息傳遞,發現肝細胞生長因子的接收器(cMet)的磷酸化和其下游數個分子的活性皆因為MUC20的表現而被抑制。由以上的實驗結果得知,MUC20在第一孕程有明顯的表現,而且對滋養層母細胞的爬行和侵襲有調控的效果,MUC20可能對於胎盤發育有重要的影響。 | zh_TW |
dc.description.abstract | Trophoblast invasion is essential for normal placental development. Lacking of hepatocyte growth factor (HGF) secretion from trophoblast causes placental defect and embryonic lethality in mice. Mucin 20 (MUC20) can interact with cMet and suppress HGF-induced signaling. In our previous study, MUC20 is expressed in human placenta. However, the expression pattern and function of MUC20 in human placenta remain largely unknown. Here, we report that MUC20 was highly expressed in human first trimester placentas by Western blot analysis. Immunohistochemistry revealed that MUC20 was mainly expressed in cytotrophoblast of cell columns and the syncytiotrophoblast of placental villi in early pregnancy, but not in decidual extravillous trophoblast (EVT). MUC20 expression in the EVT increased with gestational age. Conversely, the expression of MUC20 in syncytiotrophoblast was decreased. In trophoblast-like cells, JAR and 3A-Sub-E, overexpression of MUC20 significantly inhibited cell migration and invasion, revealed by transwell migration and matrigel invasion assays, respectively. Moreover, MUC20 overexpression suppressed HGF-induced invasion, suggesting the involvement of cMet and its downstream signaling. Our results suggest that MUC20 is differentially expressed by trophoblasts and is a novel negative regulator of trophoblast-like cell invasion throughout pregnancy. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T06:45:54Z (GMT). No. of bitstreams: 1 ntu-100-R98446007-1.pdf: 1769037 bytes, checksum: b56157c75b31413a1827298e76ff4cb4 (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | 致謝 1
中文摘要 2 Abstract 4 Chapter 1 Introduction 5 Chapter 2 Materials and Methods 11 2.1 Antibody 11 2.2 Generation of MUC20 recombinant proteins in Escherichia coli 11 2.3 Anti-MUC20N and anti-MUC20C antibody generation 13 2.4 Plasmid construction 13 2.5 Cell lines and cell culturing 14 2.6 Clinical tissue and collection 14 2.7 Overexpression of MUC20 15 2.8 Analysis of HGF-induced cell signaling 15 2.9 Western blot analysis 16 2.10 Immunohistochemistry 17 2.11 Migration and invasion assays 18 2.12 MTT assay 19 2.13 Statistical analysis 19 Chapter 3 Results 20 3.1 Generation of anti-MUC20N and MUC20C specific polyclonal antibodies. 20 3.2 Characterization of anti-MUC20 polyclonal antibodies and differential MUC20 protein expression in human placenta. 21 3.3 MUC20 had differential expression pattern in human placental villi and decidua. 23 3.4 Stable expression of MUC20 in human trophoblast-like cells 24 3.5 MUC20 overexpression suppressed the migration and invasiveness of JAR cells in serum gradient condition, but only suppresses the cell invasion in HGF gradient condition. 24 3.6 MUC20 suppressed the HGF-induced JAR cell invasion via reducing cMet activation. 26 3.7 MUC20 overexpression suppressed the migration and invasiveness of 3A-Sub-E cell in serum gradient condition, but only suppresses the cell invasion in HGF gradient condition. 27 3.8 MUC20 decreased the HGF-induced 3A-Sub-E cell invasion via down regulatory cMet activation. 28 Chapter 4 Discussion 29 Chapter 5 Reference 35 Figures 41 Figure 1 Construction of pET30a/MUC20N-His 41 Figure 2 Construction of pET30a/MUC20C-His 42 Figure 3 Construction of pSecTag2/Hygro/MUC20-myc-His 43 Figure 4 Production of soluble and insoluble MUC20C recombinant proteins 44 Figure 5 Purification of MUC20 recombinant proteins and antibodies 46 Figure 6 The specificity of MUC20N and MUC20C antibodies 48 Figure 7 The expression level of MUC20 in human placenta 49 Figure 8 Expression pattern of MUC20 in human placenta 50 Figure 9 MUC20 expression in human trophoblast-like cell lines JAR and 3A-Sub-E. 52 Figure 10 MUC20 overexpression inhibits migration and invasion of JAR cells in serum gradient condition. 53 Figure 11 MUC20 overexpression inhibits HGF-induced invasion, but not migration of JAR cells. 54 Figure 12 MUC20 suppresses HGF-induced cMet downstream signaling in JAR cell 55 Figure 13 MUC20 overexpression inhibits migration and invasion of 3A-Sub-E cells in serum gradient condition. 56 Figure 14 MUC20 overexpression inhibits HGF-induced invasion, but not in migration assay of 3A-Sub-E cells. 57 Figure 15 MUC20 suppresses HGF-induced cMet downstream signaling in 3A-Sub-E cell 58 Figure 16 The growth of JAR and 3A-Sub-E stable transfectants were not significantly affected in 48 hours by MTT assay. 59 | |
dc.language.iso | en | |
dc.title | MUC20在人類胎盤的表現與其對滋養層母細胞的影響 | zh_TW |
dc.title | Differential expression of MUC20 in Human placenta and its effect on trophoblast | en |
dc.type | Thesis | |
dc.date.schoolyear | 99-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 徐明洸(Ming-Kwang Shyu),李明學(Ming-Shyue Lee) | |
dc.subject.keyword | 黏液蛋白,胎盤,滋養層母細胞,侵襲作用,肝細胞生長因子, | zh_TW |
dc.subject.keyword | mucin,placenta,trophoblast,invasion,HGF, | en |
dc.relation.page | 59 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2011-07-25 | |
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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