新型多级循环肿瘤细胞分选和检测方法的研究

Circulating tumor cells (CTC) contains a great amount of information about cancer. The separation and analysis of CTCs have great values to the early diagnosis of cancer transfer, monitoring the recurring of cancer transfer, predicting the potential effectiveness of treatment, providing guidance to the selection of treatment plan after surgery, personized medicine as well as exploring cancer mechanism. However, the presence of CTCs in human blood usually is extremely low. A patient with transferred cancer cells in the whole blood is about 1~10 per mL. Thus, it is very challenge to have a method that can sort cells high purity, high sensitivity, high speed, and high survive rate of bioacitivity. This proposal intends to develop some new methods for cell analysis based on the extensive experience of our research group: 1) Develop new methods for the separation of cancer cells; 2）Bring gravity field into the sorting/separation of cells and explore the behavior of cells in such an environment. This may provide some theoretical insight into such a combination of gravity field and electric field; 3) Use multiorifice fractionation microchip or dielectrophoresis microfluidic chip as the preliminary cell sorting method followed by wide bore electrophoresis as the second mechanism for enhanced separation; 4) Use CCD imaging to record the whole process of cell sorting, separation, storage. It is expected that this project will result in a separation method with high recovery rate, high purity, high throughput for CTCs.

循环肿瘤细胞（CTCs）蕴含着大量的癌症信息，故CTCs的分离和检测研究对肿瘤转移的早期诊断、复发转移监控、判断预后、指导术后辅助治疗方案的选择、个体化用药及肿瘤转移机制的探索等具有重要的价值，但循环系统中CTCs的含量极低，一般癌症转移患者每毫升全血中仅有1～10个CTCs，故高纯度、高灵敏性、高速、高细胞活性的分选富集CTCs极具挑战。本项目拟结合本课题组多年大管电泳研究的经验，1）开发大管电泳分选肿瘤细胞的新方法；2）应用重力场至电泳分选细胞中，探索重力场存在下的相关电泳理论；3）以惯性迁移力微流控芯片或惯性-介电泳微流控芯片作为肿瘤细胞初选方法，将之与所开发的大管电泳分选细胞相结合，建立多级肿瘤细胞分选方法；4）通过CCD显微成像检测分选肿瘤细胞，探索分选细胞的转移、储存等后续分析。通过本项目的探索，期望开发一种高回收率、高纯度、高通量的CTCs分选方法。

近年来，肿瘤发病率逐年增高，而死亡率一直居高不下，严重威胁了人类的身心和健康。癌症的早期诊断和治疗对肿瘤的治疗和生命的延缓至关重要。本项目结合本课题组多年大管电泳研究经验，将重力场这一不利因素转化为大管电泳分离肿瘤细胞第三作用力，开发了大管电泳分选肿瘤细胞的新方法，不但实现了电泳技术分离细胞的零突破，还发现肿瘤细胞的电泳迁移行为与正常细胞确实存在差异。同时，本项目还结合微流控芯片技术，建立了多级肿瘤细胞分选方法，并探索了分选细胞的转移、裂解和内容物等后续分析，为肿瘤细胞的分选研究奠定了基础。