- 學位論文
原子力顯微鏡應用於基因轉譯細胞之特性
The Application of Atomic Force Microscopy on the Detection of Cells after Gene Transfection
摘要
在本論文中，主要將經致癌基因Aurora-A轉染之人類子宮頸癌細胞株(HeLa)培養至經由γ-APTES (3-aminopropyltriethoxysilane) 做表面化學修飾處理後的二氧化矽基板上，然後利用原子力顯微鏡於奈米尺度下來觀察細胞移除後所留在γ-APTES表面的印痕。由於細胞力學之因素，細胞會將力量施加於γ-APTES之表面上，而子宮頸癌細胞HeLa與經過致癌基因Aurora-A轉染的HeLa細胞之細胞力學必然不同。故經由原子力顯微鏡來觀察細胞移除後所留在γ-APTES表面的印痕可將細胞與基板之交互作用現象完整呈現。 我們選擇Aurora-A基因以及控制組的EGFP ( Enhanced Green Fluorescent Protein )送入子宮頸癌細胞HeLa內，並將其與控制組在和單純HeLa細胞來做比較。經由移除後之細胞印痕發現，隨著時間的增加，Hela癌細胞印痕周圍會產生深度加深的變化，而控制組表現結果與沒有做任何處理的細胞印痕結果相似。但轉染Aurora-A基因後，會使印痕深度變化有所改變，推測是因為致癌基因Aurara-A高量表現而造成細胞migration的現象產生。 利用原子力顯微鏡來觀察細胞之行為，雖然無法觀察細胞整體情形而只能單方面的對細胞周圍抓附現象進行討論，但仍不失為觀測細胞-基板間作用力的一個不錯的方法。本實驗結果或許可以提供一些關於細胞特性的訊息，使對於生物相關領域的研究人員可對細胞之變化有更進一步的了解。
並列摘要
In this thesis, attempt has been made to investigate the post-cell-removal surface morphology profiles by using atomic force microscopy for the human cervical cancer line HeLa as well as HeLa treated with the oncogene Aurora-A transfection that were cultured on 3-aminopropyltriethoxysilane ( γ-APTES ) modified silicon dioxide surface. Since force would be exerted to the γ-APTES substrate surface beneath by cells according to the cell dynamics, it is certain that the post-cell removal imprints left would be different for HeLa cell and HeLa treated with the oncogene Aurora-A transfection as the nature of the two cells are different. Therefore, the psot-cell-removal surface morphology profiles measured by atomic force microscopy can completely reflect the cell-substrate interaction property. The oncogene Aurora-A were chose for HeLa cell transfection in this work, and the EGFP ( Enhanced Green Fluorescent Protein ) was used as the control. By comparing the post-cell-removal surface morphology profiles of the cancer cell Hela, it is found that the depth along the circumference of the imprints increases over time. The transfection of EGFP into the HeLa cell does not change the cell imprint a lot and exhibits quite similar imprints as the Hela cells without any treatment. However, after transfection with the oncogene Aurora-A, the depth of the cell imprint varies prominantly. Presumably, the high expression of the oncogene Aurara-A enhances cell migration of the Hela cell and induces stronger cell-substrate interaction. . Although observing the cell behaviors by using atomic force microscopy can only detect the cell traction force along the peripheral of the imprints and is not able to determine the overall behavior of the cells, it is still a quite powerful metrology for cell-substrate interaction. The results obtained for the cell-substrate interaction can still provide some useful information for researchers in biological area.