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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/62118完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳宏文(Hungwen Chen) | |
| dc.contributor.author | Cheng-Fu Lin | en |
| dc.contributor.author | 林承賦 | zh_TW |
| dc.date.accessioned | 2021-06-16T13:28:44Z | - |
| dc.date.available | 2018-08-23 | |
| dc.date.copyright | 2013-08-23 | |
| dc.date.issued | 2013 | |
| dc.date.submitted | 2013-07-22 | |
| dc.identifier.citation | 第六章 參考文獻
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/62118 | - |
| dc.description.abstract | 摘要
細胞的移行與入侵在人類胎盤發育過程中是重要的關鍵步驟。人類滋養層細胞分化為絨毛外滋養葉細胞後會移行進入子宮內膜並入侵子宮血管管壁，藉此促進母親與胎兒養分氧氣交換循環效率之提升。先前我們確立了特定在胎盤細胞表現的轉錄因子，GCM1，會轉錄活化標的基因，HtrA4，該標的基因轉譯為一可分解細胞外基質之蛋白酶，能促進胎盤細胞之入侵能力。而最近透由染色質免疫沉澱結合晶片分析技術的分析發現一個新的GCM1標的基因，Wnt10b，該蛋白已知為WNT蛋白家族之一員。由細胞移行能力檢測實驗結果可知在BeWo細胞中阻斷Wnt10b表現將抑制其移行能力；反之如果在JAR細胞培養液中加入Wnt10b重組蛋白則可促進細胞之移行。免疫組織染色結果也顯示，在早期與晚期之胎盤組織中，Wnt10b蛋白分別出現在錨定絨毛與母親蛻膜接觸，具有移行潛力的細胞柱位置以及絨毛外滋養層細胞。更進一步，透過西方墨點法以及免疫螢光染色，可以發現Wnt10b藉由影響小GTPase蛋白家族中Rac1與Cdc42的活性，產生細胞骨架重組現象。本篇論文之實驗結果顯示GCM1藉由調控Wnt10b表現使細胞骨架產生重組，藉此調控胎盤細胞之移行。 | zh_TW |
| dc.description.abstract | Abstract
Cell migration and invasion are essential for placental development. Human cytotrophoblasts may differentiate into extravillous trophoblasts (EVTs), which migrate and invade uterus to remodel uterine arteries and enhance the maternal-fetal circulation. Our lab has demonstrated that the placental transcription factor GCM1 promotes placental cell invasion via transactivation of high-temperature requirement A4 (HtrA4) gene expression. HtrA4 is a serine protease that cleaves the extracellular matrix protein fibronectin and facilitates placental cell invasion. Our lab has recently identified Wnt10b, which is a member of WNT family protein, as a novel target gene of GCM1 by ChIP-chip analysis. By cell migration assay, we demonstrated that Wnt10b knockdown reduces the migration activity of BeWo cells, whereas Wnt10b overexpression promotes JAR cell migration. Immunohistochemistry revealed that Wnt10b protein is expressed in the cell columns of first-trimester placenta and the EVTs of full-term placenta. Furthermore, we found that Wnt10b may activate small GTPases, Rac1 and Cdc42, to induce placental cell cytoskeleton remodeling and enhance cell migration. Our study reveals a novel GCM1-Wnt10b axis in regulation of placental cell migration. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T13:28:44Z (GMT). No. of bitstreams: 1 ntu-102-R00b46027-1.pdf: 2357406 bytes, checksum: 5e15ef863b8d843b438dad3a21961922 (MD5) Previous issue date: 2013 | en |
| dc.description.tableofcontents | 目錄
目錄 i 圖表目錄 iii 中文摘要 iv 英文摘要 v 第一章 緒論 1.1 胎盤 1 1.2 GCM轉錄因子 3 1.3 WNT蛋白質家族 7 1.4 研究動機 11 第二章 材料與方法 2.1 構築重組質體 12 2.2 細胞株培養與轉染 14 2.3 慢病毒感染 15 2.4 SDS聚丙烯醯胺凝膠電泳 16 2.5 西方墨點法 16 2.6 定量及時聚合酶連鎖反應 17 2.7 Luciferase冷光報導基因活性檢測實驗 18 2.8 免疫組織染色 19 2.9 細胞移行能力檢測實驗 19 2.10 Rho蛋白家族-小GTPase活性檢測實驗 20 2.11 免疫螢光染色 20 第三章 實驗結果 3.1 GCM1轉錄因子調控胎盤細胞移行與Wnt10b基因表現 22 3.2 GCM1轉錄因子結合Wnt10b基因啟動子特定區域 22 3.3 分析Wnt10b 啟動子區域中-1.5~-2 kb片段 23 3.4 突變後之片段影響GCM1轉錄因子結合調控下游基因 24 3.5 GCM1轉錄因子在胎盤細胞調控Wnt10b表現 24 3.6 Wnt10b在胎盤組織中表現的位置 25 3.7 Wnt10b促進胎盤細胞的移行能力 26 3.8 Wnt10b活化WNT/平面細胞極化訊息路徑 26 第四章 討論與總結 29 第五章 圖表 33 第六章 參考文獻 48 圖表目錄 圖一 人類胎盤構造 33 圖二 WNT訊息路徑模式圖 34 圖三 GCM1轉錄因子調控胎盤細胞移行與Wnt10b基因表現 35 圖四 GCM1轉錄因子結合Wnt10b基因啟動子特定區域 37 圖五 分析Wnt10b啟動子區域中-1.5~-2 kb片段 38 圖六 突變後之片段影響GCM1轉錄因子結合調控下游基因 39 圖七 GCM1轉錄因子在胎盤細胞調控Wnt10b表現 40 圖八 Wnt10b在胎盤組織中表現的位置 41 圖九 Wnt10b促進胎盤細胞的移行能力 42 圖十 Wnt10b在胎盤細胞中不顯著影響典型WNT訊息路徑 43 圖十一 Wnt10b活化WNT/平面細胞極化訊息路徑 45 圖十二 GCM1調控Wnt10b進行平面細胞極化路徑促進細胞移行模式圖 47 | |
| dc.language.iso | zh-TW | |
| dc.subject | 胎盤 | zh_TW |
| dc.subject | 細胞骨架重組 | zh_TW |
| dc.subject | GCM1 | zh_TW |
| dc.subject | Wnt10b | zh_TW |
| dc.subject | Rac1/Cdc42 | zh_TW |
| dc.subject | 細胞移行 | zh_TW |
| dc.subject | cytoskeleton remodel | en |
| dc.subject | GCM1 | en |
| dc.subject | Rac1/Cdc42 | en |
| dc.subject | placenta | en |
| dc.subject | cell migration | en |
| dc.subject | Wnt10b | en |
| dc.title | Wnt10b作為GCM1轉錄因子之標的基因透過WNT/平面細胞極化訊息路徑調控胎盤細胞移行 | zh_TW |
| dc.title | Wnt10b, a novel target gene of GCM1, regulates placental cell migration through the WNT/PCP pathway | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 101-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 張震東(Geen-Dong Chang),李明亭(Ming-Ting Lee),張茂山(Mau-Sun Chang),張功耀 | |
| dc.subject.keyword | Wnt10b,GCM1,Rac1/Cdc42,胎盤,細胞移行,細胞骨架重組, | zh_TW |
| dc.subject.keyword | Wnt10b,GCM1,Rac1/Cdc42,placenta,cell migration,cytoskeleton remodel, | en |
| dc.relation.page | 52 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2013-07-22 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 生化科學研究所 | zh_TW |
| 顯示於系所單位： | 生化科學研究所 | |
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