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

Wnt10b, a novel target gene of GCM1, regulates placental cell migration through the WNT/PCP pathway

林承賦(Cheng-Fu Lin)

指導教授 ： 陳宏文

國立臺灣大學/生命科學院/生化科學研究所/碩士(2013年)

https://doi.org/10.6342/NTU.2013.10950

摘要

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。

關鍵字

Wnt10b ； GCM1 ； Rac1/Cdc42 ； 胎盤 ； 細胞移行 ； 細胞骨架重組

並列摘要

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.

並列關鍵字

Wnt10b ； GCM1 ； Rac1/Cdc42 ； placenta ； cell migration ； cytoskeleton remodel

參考文獻

第六章 參考文獻

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