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Title: | 探討上皮-間質轉化與細胞於幾丁聚醣之脫附能力關係 The Relationship between Epithelial-Mesenchymal Transition and Cell Detachment Ability on pH-responsive Chitosan |
Authors: | Meng-Chi Hsieh 謝孟其 |
Advisor: | 楊台鴻 |
Keyword: | 幾丁聚醣,酸鹼敏感,轉化生長因子β1,上皮-間質轉化,整聯蛋白β4,細胞脫附, Chitosan,pH-responsive,TGF-β1,EMT,Integrin β4,Cell detachment, |
Publication Year : | 2015 |
Degree: | 碩士 |
Abstract: | 近年來,細胞操縱的技術漸趨發達並被開發及應用於多方面的研究,例如利用光學或是電磁特性的差異來分離不同種類的細胞甚至控制細胞的行為模式,而酸鹼敏感幾丁聚醣在近幾年則被發現可以透過改變環境培養液的酸鹼值達到調控細胞貼、脫附的效果;另一方面,上皮-間質細胞轉化的現象已經被視為在癌症生長及移轉過程中至關重要的步驟,而其機制也被深入研究並探討其中關鍵的影響因子。因此在本研究中,我們利用轉化生長因子β1來誘導上皮細胞進行上皮-間質細胞轉化,並以表現上皮和間質細胞特性的生物標記比較轉化的程度;再者,為了觀察細胞脫附率的變化,我們亦將細胞種於酸鹼敏感幾丁聚醣上並進行細胞脫附實驗,同時檢視細胞貼附的調停分子-整聯蛋白β4,藉此驗證其在細胞移動上之角色;另外,也進行細胞分選實驗,期許能建立癌症轉移的模型。我們發現經過轉化的上皮細胞確實從原本較圓以及群聚的型態轉變成細長且偏向分散的情形,而生物標記的表現也有明顯的消長:間質細胞特有的波形蛋白表現增強;反之,屬於上皮細胞的鈣黏蛋白表現則減弱。此外,在細胞脫附率方面,隨著轉化生長因子β1濃度的提高而逐漸增加,整聯蛋白β4的表現則下降,這些結果再次顯示轉化生長因子β1確實能引發上皮-間質轉化的發生,而轉化後的細胞其移動能力則明顯增加。 pH-responsive chitosan, a kind of smart polymers, has been found to be able to control cell adhesive behavior by changing the pH value of environmental medium. Meanwhile, epithelial-mesenchymal transition (EMT), which is regarded as the vital phenomenon in cancer progression and metastasis, has also been studied to understand its signaling mechanism and discover the factors that influence EMT. In this study, we utilized transforming growth factor beta 1 (TGF-β1) to induce EMT on A549, PK15 and hRPTEC cells and then biomarkers of epithelial and mesenchymal cells, E-cadherin and vimentin, were observed to compare the degree of EMT. To check the change of cell detachment ratio, three cells and their alterative types were executed by cell detachment assay on pH-responsive chitosan. Furthermore, a cell adhesion mediator, integrin β4 (ITGb4), was examined to validate the role in cell mobility and migration. We found that the transformation of cell morphologies from more round shape and collective type to elongated form and individual style during the EMT, which was accompanied with the up-regulation of vimentin and down-regulation of E-cadherin. Besides, cell detachment ratio increased as the concentration of TGF-β1 rose up, which was consistent with the decreased expression of ITGb4. The results indicated that TGF-β1 did cause the occurrence of EMT and after EMT occurred, cell migratory ability enhanced obviously. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/53767 |
Fulltext Rights: | 有償授權 |
Appears in Collections: | 醫學工程學研究所 |
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ntu-104-1.pdf Restricted Access | 2.78 MB | Adobe PDF |
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