The Evolvement of the Cell-Substrate Interaction over Time for Normal Cells and Cancer Cells and for Cancer Cells Treated with Anti-Cancer Medicine

• Main Authors: Eric Lee, 李文淵
• Other Authors: 吳幼麟
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/38955474704779877017

碩士 === 國立暨南國際大學 === 生物醫學科技研究所 === 98 === In this paper, we discussed the cell-substrate interaction by adding several kinds of stimulus to cancer cells and normal cells. In the work, we seeded the living cells onto the silicon wafer surface that was modified with 3- aminopropyltriethoxysilane (γ-APTES)for different periods of time, then removed the cells from the chip and, measured the surface morphology profiles of the γ-APTES layer by atomic force microscopy (AFM) to realize the possible cell-substrate interactions between the cell and the substrate beneath, and to observe the evolvement of cell imprints over time. The silicon wafer which 2nm-thick SiO2 on the surface was divided into several pieces, the area of each is about 1cm2. The APTES was then coated onto the surface by spin coating process at 3000 rpm, 30s followed by 5 min baking at 120 ℃. All the samples were then put in to a glass container and subjected to a high-temperature and high pressure sterilization process to as sure that no bacteria inflection of the samples. In this work, the human lung adenocarcinoma cells, H1299 and A549, and Madin-Darby Canine Kidney epidermal cell (MDCK),were cultivated first, in an incubator at 37 ℃ and 5 % CO2 environment, and subculturing was carried out for every two days’ cultivation. After 1, 4, 12, 24 and 72 hr cultivation of the cells on γ-APTES surface, we removed the cells and measured the surface morphology images and profiles of the imprints left on the surface. From the experimental data, we found that the depth of the cell imprints were dependent on the cultivation time. The longer the cultivation time is the deeper the imprint depth will be. However, the imprint depth for the cell after 24 hr cultivation is the same as that after 72 hr cultivation. The difference of the cell imprints between the normal cells (MDCK) and cancer cells (H1299, A549) is that the cancer cells exhibit a deeper trench on one side of the imprints, and the force at the central region of the cell begins to prevail after several hours of cultivation. On the contrary the force of the normal cells seems to be applied to the substrate evenly which causes the cell imprint flat over the whole cell-substrate adhesion region. In this work, two anti-cancer medicines, staurosporine and Iressa, were added into the cell cultivation solution to see how they affected the cell-substrate interaction. We tried to investigate and explain the role play of these anti-cancer medicines in cell-substrate interaction from the cell molecular biology point of view. The surface morphology profiles of the cells added with different concentration of staurosporine, 10-1, 1, 10, and 102 nM, were measured, which show clearly that the cell imprints are severely influenced by the concentration changes. We noticed that the higher the staurosporine concentration was the shallow the depth of the imprint would be. Cell recovery experiment was also conducted by substitute the staurosporine-contained medium after 24 hr cell cultivation with fresh medium, we found that the final cell imprint of the cell after another 24 hr cultivation was the same as that not being treated with staurosporine. For another anti-cancer medicine Iressa, which has been used particularly to inhibit the epidermal growth factor receptor (EGFR), we found that the cell imprints were not affected by the concentration of Iressa. In this thesis, a novel method using the post-cell-removal surface morphology profiles measured by AFM was proposed for the investigation of ll-substrate interaction. By adding the cancer cells with anti-medicines, we verified the proposed method is a promising one for cell traction force as well as cell behavior observation.