新型纳米微流控芯片在胰腺癌CTC检测中的模拟与优化

The simulation and optimization of new nano-microflluidic chip in the CTC detection of pancreatic cancer

目的探索新型纳米微流控芯片(Nextctc)分离循环肿瘤细胞(circulating tumor cells,CTC)的实验条件,探讨该技术用于胰腺癌CTC捕获的可行性。方法采用肿瘤细胞株模拟CTC,以不同进样速度进样,获得Nextctc最佳进样速度并计算捕获效率,并用胰腺癌细胞株模拟CTC加以验证;同时采集晚期胰腺癌外周血送检CellSearch及Nextctc,比较两者在胰腺癌CTC捕获中的效率。结果在1.0、1.5、2.0mL/min进样速度下Nextctc的平均富集率分别为57.7%、66.3%、75.2%,继续升高进样速度分离系统出现堵塞,该结果在胰腺癌细胞株模拟CTC实验中得到验证,综合富集效率及通畅性确定2.0 mL/min为最佳进样速度;采用Cellsearch法与Nextctc法同时检测20例胰腺癌患者CTC其捕获阳性率分别20%及100%,两组方法存在统计学差异(P <0.05)。结论 Nextctc纳米微流控芯片在CTC分离捕获中具有高通量、高富集率等特点,是新型的CTC捕获平台,可用于胰腺癌CTC的检测研究。

objective To explore the experiment conditions under which a new nano-microfluidic chip （Nextctc） was used to separate circulating tumor cells （CTC）, and to probe into the feasibility of capturing CTC of pancreatic cancer with this technology. Methods Tumor cell strain was adopted to simulate CTC. Samples were injected at different speeds. The best sample injection speed of Nextctc was acquired and the capture rate was calculated. Pancreatic cancer cell strain was used to simulate CTC for further verifcation. In the meantime, the peripheral blood of advanced pancreatic cancer was collected and submitted to CellSearch and Nextctc, in an effort to compare the efficiency of both methods in capturing the CTC of pancreatic cancer. Results The mean enrichment ratio of Nextctc at the injection speed of 1.0 mL/min, 1.5 mL/min, and 2.0 mL/min was 57.7%, 66.3%, and 75.2% respectively. Blocking occurred in the separation system when the injection speed was increasing unceasingly. The result had been verifed in the experiment in which pancreatic cancer was used to simulate CTC. 2.0 mL/min was confrmed as the best injection speed, according to the enrichment effciency and liquidity. The CTC of 20 cases of pancreatic cancer was detected with Cell Search and Nextctc, and the capture positive rate was 20% and 100% respectively. There was signifcant difference between two groups （P〈0.05）. Conclusion Nextctc nano-microfuidic chip is characterized by high throughput, high enrichment ratio in the separation and capture of CTC. It is a new type of CTC capture platform, and which can be used in the detection and study of CTC of pancreatic cancer.