中文摘要
耐药性是导致肺癌化疗失败的主要原因之一，多种耐药蛋白参与该过程。目前常用的Western-blot, RT-PCR及MTT等耐药性分析技术不仅操作复杂、灵敏度较低，而且难以实现多种耐药蛋白的联合检测，特别是与体内微环境相差甚远,检测结果与临床相符率不高，因而应用受到了一定的限制。微流控芯片是一项发展中的技术，以高通量和规模集成为主要特点，可以模拟体内的生理环境，进行生理或近生理条件下的细胞功能性研究。本项目通过微阀微泵连接，构建流动介质微流控芯片体系，模拟体内微环境调整培养基、药物等流速，分别对四种人肺癌标准细胞株中P-gp, MRP, LRP, GST-π 和TopoIIα的表达以及它们对VP-16、顺铂等化疗药物的耐药性进行了检测，分析耐药蛋白与肺癌耐药的相关性。作为对照，同时完成了静态条件下的细胞培养、蛋白检测及MTT分析等对比实验。结果表明，微流控芯片体系肺癌耐药性研究结果与临床常用技术结果一致，但微流控芯片体系显示了细胞生长状态好，与体内生理环境更接近以及试剂消耗量少、反应速度快、检测效率高等优点，表明该体系可用于细胞培养及功能性研究，在医学上具有良好的应用前景。
英文摘要
Chemotherapy resistance is the main problem of lung cancer. Many drug resistance related proteins are involved in the process. The common used drug resistance analysis techniques such asWestern-blot, RT-PCR and MTT have the lower coincident with the clinical because of the complex operation and the lower sensitivity detection in vitro, which are far away from the microenvironment in vivo. The microfluidics chip system is a promising technique with the ability to stimulate the microenvironment in vivo and can be used in the study under the physiological or near physiological situation with the advantages of high-throughput and integration. This project aimed at the analysis of the relationship between the expression of P-gp, MRP, LRP, GST-? , TopoII?and the resistance to chemotherapy to VP-16，cisplatin and other drugs on four standard cell lines of human lung carcinoma mimicking the environment in vivo on micro fluidic system with a flow mudium by controlling the speed of culture medium and drug concentration referring to the blood stream speed and drug metabolize mode in vivo with the help of micro-valve and micro-pump. In contrast the clinical current used methods of dish cell culture and MTT drug resistance analysis were performed in parallel on thse cell lines as well. The results indicated that there was correlation between the expression of P-gp, MRP, LRP, GST-? ,TopoII? and the resistance to those anti-cancer drugs. Moreover, the results from the microfluidic chip system was similar to that with the traditional methods but the former showed many advantages including lower samples consumption, rapid analyses and high efficiency, especially the simulation to the environment in vivo with the good growth condition of the cells. All these mean the microfluidic chip system with flow mudium can be used in the cell culture and the analysis of the cell function with great prospect in medicine.
结题摘要
耐药性是导致肺癌化疗失败的主要原因之一，多种耐药蛋白参与该过程。目前常用的Western-blot, RT-PCR及MTT等耐药性分析技术不仅操作复杂、灵敏度较低，而且难以实现多种耐药蛋白的联合检测，特别是与体内微环境相差甚远,检测结果与临床相符率不高，因而应用受到了一定的限制。微流控芯片是一项发展中的技术，以高通量和规模集成为主要特点，可以模拟体内的生理环境，进行生理或近生理条件下的细胞功能性研究。本项目通过微阀微泵连接，构建流动介质微流控芯片体系，模拟体内微环境调整培养基、药物等流速，分别对四种人肺癌标准细胞株中P-gp, MRP, LRP, GST-π 和TopoIIα的表达以及它们对VP-16、顺铂等化疗药物的耐药性进行了检测，分析耐药蛋白与肺癌耐药的相关性。作为对照，同时完成了静态条件下的细胞培养、蛋白检测及MTT分析等对比实验。结果表明，微流控芯片体系肺癌耐药性研究结果与临床常用技术结果一致，但微流控芯片体系显示了细胞生长状态好，与体内生理环境更接近以及试剂消耗量少、反应速度快、检测效率高等优点，表明该体系可用于细胞培养及功能性研究，在医学上具有良好的应用前景。