【作者】 郑波；

【导师】 曾苏；

【作者基本信息】 浙江大学 ， 药学， 2012， 硕士

【摘要】 恶性肿瘤是一种严重威胁人类健康的疾病,并且其发病率近年来一直都处于逐年上升的趋势。目前,化学药物治疗仍是肿瘤的重要治疗手段。因此,如何研发出能够有效抑制癌细胞生长增殖,或对其直接具有杀伤性作用的预防或治疗性药物,是肿瘤防治的重要环节。随着医药技术的快速发展,体外培养的肿瘤细胞系,为抗肿瘤药物的筛选与药效评价提供了一个有效的生物学方法。与此同时,随着微纳米技术的快速发展,近年来兴起了采用微纳米电子传感器芯片对细胞或组织进行直接测量的研究。基于细胞电阻抗传感检测的ECIS (electrical cell-substrate impedance sensor)技术,便是一种能够同时测量多组不同细胞的电阻变化、膜电容变化、以及细胞层-基底膜空间变化的细胞生理与病理研究的细胞传感器(cell based biosensor)技术。通过微安级的电流测量,可以实时连续的量化研究细胞外基质与细胞增殖之间的相互作用,测量贴壁细胞迁移过程中细胞形态变化。本研究在对细胞与电极界面阻抗检测理论分析的基础上,采用微纳米电子技术制备了以交错方式间隔排列的叉指电极阵列(interdigital electrode arrays)传感器芯片,搭建了阻抗检测分析系统,以人参皂苷Rh2等抗癌药物对人肺腺癌A549细胞模型的作用研究为例,评价了人参皂苷Rh2对肿瘤细胞的作用。并同时采用计算机辅助分子对接和药效团模型技术,对Bc1-2、Caspase-3、CKD2、PKC等细胞凋亡相关蛋白与Rh2的细胞凋亡途径进行了研究。探讨了将计算机辅助药物设计方法与生物传感器技术有效结合,对人参皂苷Rh2的抗癌作用机理进行深入研究的可行性。总之,本文以人参皂苷Rh2等抗癌药物对人肺腺癌A549细胞模型的作用研究为例,结合计算机辅助分子对接和药效团模型技术,建立了能对抗肿瘤药物进行实时、无损分析的细胞微电子传感器芯片技术方法。更多还原

【Abstract】 Malignant tumor is a serious threat to human health and disease. The number of new cases increased greatly in recent years. Now, chemotherapy is still one of successful therapeutic methods to malignant tumors. Therefore, how to find inhibitors to grows and proliferations of cancer cells, and find drugs in prevention and treatment affection with abilities of killing cancer cells directly, are all very important issues for tumors therapy. With the quickly development of biomedical techniques, in vitro cultured cancer cell lines have become available bioassays for screening and evaluating anti-cancer drugs. At the same time, with the development of micro- and nano- technology, lots of micro- and nano-sensors have been used to measure tissue or cells directly. Electrical cell-substrate impedance sensor (ECIS) is just one of those kinds of cell-based biosensors, which can be used to detect cells physiological or pathological changes, such as cell impedance, membrane capacitance, and spatial characters in cell-substrate interface in different culture channels. The sensor can monitor interactions of cell proliferations to extracellular matrix, and the morphological changes of the attached cells, in a real-time and quantitative way by microampere current measurement.Based on analysis of the cell-substrate impedance models of the theoretic analysis, interdigital electrode arrays were fabricated by micro- electromechanial system (MEMS) in this paper. The sensor was equipped with sample giving system and control system. Anti-cancer drugs, such as ginseng saponin Rh2, were analyzed by applied it to cancer cells of human lung adenocarcinoma cell line A549 cultured on the electrodes. At the same time, methods of computer aided drug design of ligand and structure based pharmacophore modeling and molecular docking, was both used to analyze the drug induced apoptosis by effacing on cell apoptosis-related proteins, such as Bcl-2, Caspase-3, CKD2, and PKC. After all, the combine of the biomedical sensors with computer aided drug design was discussed as a potential plant form for deeply understanding the mechanisms of anti-cancer drugs such as ginseng saponin Rh2.Anyway, ginseng saponin Rh2 was applied to cancer cells of human lung adenocarcinoma cell line A549 in this research. Integrated with ligand and structure based pharmacophore modeling and molecular docking, a microelectronic sensor is designed, and it provides a viable real time and non-invasive platform for anti-cancer drugs screening and analysis.更多还原