以氧化鋅奈米線指叉電極晶片檢測癌細胞分泌之外泌體

外泌體能夠被體內各種細胞分泌，包括腫瘤細胞、神經元和免疫細胞。更重要的是，外泌體涉及腫瘤抗原、抗腫瘤免疫反應和腫瘤診斷，且其富含蛋白質的標記物，其中包括CD9、CD63和CD81。越來越多的證據表明外泌體通過調節腫瘤微環境而參與腫瘤的遷移，因此癌症衍生的外泌體成為具有吸引力的非侵入性診斷和預後生物標記物。在本研究中，使用氧化鋅奈米線基於指叉電極方式檢測癌細胞分泌之外泌體。在定量外泌體之前，首先以掃描式電子顯微鏡、能量散射X射線光譜儀和X 光繞射光譜儀對氧化鋅奈米線進行物性分析；再來使用Nanosight粒徑分析儀與掃描式電子顯微鏡確認外泌體之濃度、尺寸與表面型態，接著以APTES和GA對氧化鋅奈米線進行改質，並在氧化鋅奈米線上鍵結抗體對其生物功能化，使其具有鍵結外泌體之能力，最後以電性方式與阻抗方式對SAS口腔癌和A549肺癌細胞分泌之外泌體進行線性濃度量測。兩種量測方式下，在稀釋倍率範圍為2000X到10000X內皆有良好的線性趨勢。人體中可能同時存在兩種癌細胞，因此得知癌細胞之濃度比以利醫護人員判斷是一件相當重要的事情。在本論文中使用量測線性濃度所得之校正曲線對不同濃度之混合外泌體進行驗證，當兩種外泌體濃度分別為3：7 ； 5：5 ； 7：3時，計算後所得結果分別為2.6：7.4 ； 4.7：5.3 ； 6.1：3.9，實驗濃度與計算濃度兩者間差異不大，由此證明驗證結果之成功。最後，干擾性實驗也說明本論文之外泌體感測器對於干擾物之選擇性與抗干擾性具有一定能力。

關鍵字

電性量測；阻抗量測；氧化鋅奈米線；外泌體

並列摘要

Exosomes can be secreted by various cells in the human body, including tumor cells, neurons, and immune cells. More importantly, exosomes involved in tumor antigens, anti-tumor immune reactions, and tumor diagnosis. On the other hands, the surface of exosomes has wealth of protein markers including CD9, CD63, and CD81. Growing evidences showed the participation of exosomes in tumor migration and invasion through the regulation of the tumor microenvironment, therefore, cancer-derived exosomes had become attractive non-invasive diagnostic and prognostic biomarkers. In this study, the exosomes that secreted by cancer cells are detected by interdigitated electrode with zinc oxide nanowires. Before quantifing the amount of exosomes, first, the physical properties of zinc oxide nanowires are analyzed by SEM, EDX, XRD; next, the concentration, size and morphology of the exosomes are characterized by NTA and SEM, next, using APTES and GA to modify the zinc oxide nanowires, following using antibody to biofunctionalized the zinc oxide nanowires. When the above-mentioned is completed, the biofunctionalisation of zinc oxide nanowires is allowed it to bind the exosomes, on the other hands, the measuring of linear concentrations of the SAS cell (Oral cancer) and A549 cell (Lung cancer) secretory exosomes was performed by electrical and impedance methods. Under the electrical and impedance methods, there is a good linear trend in the range of dilution rate from 2000X to 10000X. However, there are maybe two kinds of cancer cells in the human body at the same time, as a result, it is very important to know the concentration ratio between the cancer cells so that heiping medical personnel to make a decision which treating what kinds of the cancer cell at first. In this paper, we verify the concentration ratio between two exosomes by using calibration curve which obtained by the linear concentration. When the concentrations of the two exosomes were 3:7 ; 5:5 ; 7:3, the results of calculation were 2.6:7.4; 4.7:5.3; 6.1: 3.9, and we can find the difference between the result of the experiment and the calculation is small, which proved the success of the results. Finally, the interfering test also showed a strong ability of the sensors to against the interference.

並列關鍵字

Electrical measurement ； Impedance measurement ； Zinc oxide nanowires ； Exosomes

參考文獻

[1] K.D. Miller, R.L. Siegel, C.C. Lin, A.B. Mariotto, J.L. Kramer, J.H. Rowland, K.D. Stein, R. Alteri, and A. Jemal,“Cancer treatment and survivorship statistics,” CA Cancer J. Clin., vol. 66, pp. 271-289, 2016.

Google Scholar

[2] C.A. Lieber, S.K. Majumder, D.L. Ellis, D.D. Billheimer, and A. Mahadevan‐Jansen,“In vivo nonmelanoma skin cancer diagnosis using Raman microspectroscopy,” Lasers Surg. Med., vol. 40, pp. 461-467, 2008.