基于纳米纤维材料的循环肿瘤细胞高效捕获与释放研究

Malignancies are a type of fatal diseases with serious threat to human health, which has become one of main causes of death for adults in China. There is a considerable body of evidence indicating that circulating tumor cells (CTCs) can travel to different tissues of the body, which is regarded as the cellular origin of metastases. In early stage metastasis, it is difficult to identify the metastatic sites for biopsy. CTCs can be regarded as the "liquid biopsy" of the tumor, thus providing a earlier and more safe access to potentially fatal metastases of tumor. However, detection and characterization of CTCs, as well as following manipulation and analysis of single CTC, have been technically challenging due to the extremely low amount ( few to several hundreds cells/mL) in the blood. This project is based on the previous work that CTCs have been successfully captured from blood samples of gastric and colorectal cancer patients by using electrospun TiO2 nanofiber structured substrates. Further research on CTCs will be expanded in MnO2 nanofibers-based CTC-capture microfluidic-device which can overcome the shortcoming of these current capture-devices that disable controllable release of captured CTCs, and improvement of CTC-capture efficiency and simplification of manipulation process. Successful implementation of this project will expand CTC researches and provide powerful tools and valuable information for the following research areas, such as early cancer diagnosis, estimation of anticancer drug therapeutic effect, point of care treatment, analysis of radio- and chemo-therapeutic effect, new anti-cancer drug screening.

恶性肿瘤是一类严重威胁人类生命健康的致命疾病，是中国成年人主要死因之一。肿瘤扩散复发是原发肿瘤不断转移的过程。循环肿瘤细胞（CTC）能扩散到人体不同的部位。 活检很难识别早期转移。CTC可作为肿瘤的液检标本，从而为潜在致命转移提供更早期和安全的检测渠道。CTC数量非常稀少(每毫升血几个到数百个)，因此捕获病人外周血中的CTC进而深入开展单肿瘤细胞的操控和分析是一项艰巨的挑战。 本项目将基于前期工作（用纳米纤维衬底成功实现胃癌和结直肠癌病人血样中CTC的捕获），接下来将开展基于MnO2纳米纤维衬的微流控型捕获芯片以克服现有捕获芯片无法对已捕获的CTC进行可控释放的缺点，并进一步提高捕获效率和简化操控流程。 本项目的成功实施将会拓展循环肿瘤细胞的研究，并将为癌症早期诊断、抗肿瘤药物药效评价、放化疗疗效分析、癌症的个体化治疗，新型抗癌药物筛选等研究提供有力手段和有价值的信息。

恶性肿瘤是一类严重威胁人类生命健康的致命疾病，是中国成年人主要死因之一。肿瘤扩散复发是原发肿瘤不断转移的过程。循环肿瘤细胞（CTC）能扩散到人体不同的部位。活检很难识别早期转移。CTC可作为肿瘤的液检标本，从而为潜在致命转移提供更早期和安全的检测渠道。CTC数量非常稀少(每毫升血几个到数百个)，因此捕获病人外周血中的CTC进而深入开展单肿瘤细胞的操控和分析是一项艰巨的挑战。我们针对以上问题进行了探索和研究，首先我们提出了一种新型的PMMA芯片的制备方法，该方法利用激光消融术在PMMA基质材料上制备出沟道结构，然后利用OCA膜和LOCA胶对PMMA芯片进行低温键合，其优点是键合效率和强度均得到较大提高，以确保实验中无漏液发生，其成果发表在2016年的Microsystem Technologies杂志上；与此同时，我们发展了一种新型的流速测量技术，实现了对微流控芯片的微流体（如人体外周血）精确且实时的流速监测，相关成果正在编写论文；然后，我们提出了这种基于斜面芯片的循环肿瘤细胞的高效的捕获方法，该方法利用肿瘤细胞和血液细胞在尺寸和核质比上的差异，进行CTC的高效且快速分离，并且该方法不依赖于抗原－抗体反应，能够捕获EpCAM低表达的CTCs，并且我们验证了该方法在肝癌、乳腺癌、肺癌和胃癌等病人血样CTC检测研究中的实用性和优势，相关成果正在编写论文；最后，我们提出了一种新型的CTCs高效释放方法：在斜面芯片衬底上低温合成ZnO纳米线阵列，完成CTC捕获以后，利用稀磷酸溶液溶解ZnO纳米线衬底以实现批量的CTC的高效的释放，并验证了其释放的细胞具有较高的生物活性，相关研究成果正在编写论文。