Chelakkot Chaithanya, Ryu Jiyeon, Kim Mi Young, Kim Jin-Soo, Kim Dohyeong, Hwang Juhyun, Park Sung Hoon, Ko Seok Bum, Park Jeong Won, Jung Moon Youn, Kim Ryong Nam, Song Kyoung, Kim Yu Jin, Choi Yoon-La, Lee Hun Seok, Shin Young Kee
Technical Research Center, Genobio Corp., Seoul 08394, Korea.
Department of Internal Medicine, Seoul National University Boramae Medical Center, Seoul 07061, Korea.
Micromachines (Basel). 2020 May 30;11(6):560. doi: 10.3390/mi11060560.
Here, we validated the clinical utility of our previously developed microfluidic device, GenoCTC, which is based on bottom magnetophoresis, for the isolation of circulating tumor cells (CTCs) from patient whole blood. GenoCTC allowed 90% purity, 77% separation rate, and 80% recovery of circulating tumor cells at a 90 μL/min flow rate when tested on blood spiked with epithelial cell adhesion molecule (EpCAM)-positive Michigan Cancer Foundation-7 (MCF7) cells. Clinical studies were performed using blood samples from non-small cell lung cancer (NSCLC) patients. Varying numbers (2 to 114) of CTCs were found in each NSCLC patient, and serial assessment of CTCs showed that the CTC count correlated with the clinical progression of the disease. The applicability of GenoCTC to different cell surface biomarkers was also validated in a cholangiocarcinoma patient using anti-EPCAM, anti-vimentin, or anti-tyrosine protein kinase MET (c-MET) antibodies. After EPCAM-, vimentin-, or c-MET-positive cells were isolated, CTCs were identified and enumerated by immunocytochemistry using anti-cytokeratin 18 (CK18) and anti-CD45 antibodies. Furthermore, we checked the protein expression of PDL1 and c-MET in CTCs. A study in a cholangiocarcinoma patient showed that the number of CTCs varied depending on the biomarker used, indicating the importance of using multiple biomarkers for CTC isolation and enumeration.
在此，我们验证了我们之前开发的基于底部磁泳的微流控装置GenoCTC从患者全血中分离循环肿瘤细胞（CTC）的临床实用性。当在添加了上皮细胞粘附分子（EpCAM）阳性的密歇根癌症基金会-7（MCF7）细胞的血液上进行测试时，GenoCTC在流速为90μL/分钟的情况下，可实现循环肿瘤细胞90%的纯度、77%的分离率和80%的回收率。使用非小细胞肺癌（NSCLC）患者的血样进行了临床研究。在每位NSCLC患者中发现了数量不等（2至114个）的CTC，对CTC的连续评估表明，CTC计数与疾病的临床进展相关。在一名胆管癌患者中，还使用抗EpCAM、抗波形蛋白或抗酪氨酸蛋白激酶MET（c-MET）抗体验证了GenoCTC对不同细胞表面生物标志物的适用性。在分离出EpCAM、波形蛋白或c-MET阳性细胞后，使用抗细胞角蛋白18（CK18）和抗CD45抗体通过免疫细胞化学对CTC进行鉴定和计数。此外，我们检查了CTC中PDL1和c-MET的蛋白表达。一项针对胆管癌患者的研究表明，CTC的数量因所使用的生物标志物而异，这表明使用多种生物标志物进行CTC分离和计数的重要性。
