An Investigation of A549 Human Lung Cancer Cells Treated with Colchicine Using Fourier Transform Infrared Spectra

利用傅立葉轉換紅外線顯微光譜儀探討瞭解秋水仙素與A549人類肺癌細胞間之細胞內交互作用

鄭奕帝(Yih-Dih Cheng) ； 徐美華(Mei-Hua Hsu) ； 陳俊仁(Chun-Jen Chen) ； 謝明杰(Ming-Chieh Hsieh) ； 林智廣(Chih-Kuang Lin) ； 賴雅韻(Ya-Yun Lai)

《中山醫學雜誌》 21卷4期 (2010/12) Pp. 341-353

https://doi.org/10.30096/CSMJ.201012.0002

摘要

為了瞭解秋水仙素(colchicine)與A549人類肺癌細胞間之細胞內交互作用的情形，我們利用了傅立葉轉換紅外線(FTIR)顯微光譜儀結合二次微分分析技術獲得A549細胞的IR圖譜與確認IR圖譜的波峰位置及波峰解析。當A549細胞經由秋水仙素作用後，其脂質的含量隨即減少，而此可由位在脂質吸收區之2962cm^(-1)，2916cm^(-1)，2875cm^(-1)及2847cm^(-1)的波峰位置改變而證實之。A549細胞經由秋水仙素作用後，A549細胞的醯胺鍵Ⅰ(1650cm^(-1))及醯胺鍵Ⅱ(1540cm^(-1))之α-helix波峰位置被位移至高頻率吸收區。隨著秋水仙素濃度的增加，核苷酸PO2(上標 -)伸縮作用吸收峰位置1087cm^(-1)及1240cm^(-1)亦可被觀察到明顯的不同改變。A549細胞進行細胞有絲分裂作用間的差異是很明顯的，特別是在細胞蛋白質構型的改變。這些結果指出A549細胞的α-helix結構的構型變化在分裂的細胞蛋白質中隨著秋水仙素濃度增加而些微地被誘導。而本研究中A549細胞的細胞蛋白質結構變化情形與先前的藥物與細胞間交互作用模式之研究報告相類似。

關鍵字

人類肺癌細胞A549 ； 秋水仙素 ； 二次微分分析 ； 蛋白質二次結構 ； 傅立葉轉換紅外線

並列摘要

In this study, we used Fourier Transform Infrared (FT-IR) spectroscopy with second-derivative analysis techniques to obtain Infrared (IR) spectra of A549 cells and confirm the peak positions and assignments of the IR spectra to characterize intracellular protein secondary structure of A549 human lung cancer cells treated with colchicine. Treatment with colchinine reduced the lipid content decreased in A549 cells, with peak shifts occurring at 2962, 2916, 2875, and 2847 cm^(-1). After treatment with colchicine, the peak positions of α-helix of amide Ⅰ (1650 cm^(-1)) and amide Ⅱ (1540 cm^(-1)) in A549 cells were shifted to high frequency. The differences between A549 cells that underwent mitosis were significant, especially with regard to the conformational changes of the proteins. We also observed that increasing concentrations of colchicine led to clear differences in nucleic acid PO2(superscript -) stretching (1087 and 1240 cm^(-1)). These results suggest colchicine dose-dependently induced conformational changes of a-helical secondary structures in A549 cells. However, the band at 965 cm^(-1) did not show distinct changes, indicating no marked changes of this DNA stretching vibration. Therefore, these changes of the protein structures may be related to the mode through which the agents interact with the cell, as has been reported previously.

並列關鍵字

A549 human lung cancer cells ； colchicine ； second-derivative analysis ； protein secondary structure ； FT-IR