Lv Song-Wei, Wang Jing, Xie Min, Lu Ning-Ning, Li Zhen, Yan Xue-Wei, Cai Si-Liang, Zhang Ping-An, Dong Wei-Guo, Huang Wei-Hua
Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education) , College of Chemistry and Molecular Sciences , Wuhan University , Wuhan 430072 , China . Email:
Department of Gastroenterology , Renmin Hospital of Wuhan University , Wuhan 430060 , China . Email:
Chem Sci. 2015 Nov 13;6(11):6432-6438. doi: 10.1039/c5sc01380a. Epub 2015 Jul 30.
Isolation, release and culture of rare circulating tumor cells (CTCs) may, if implemented, promote the progress of individualized anti-tumor therapies. To realize the release of CTCs without disruption of their viability for further culture and analysis, we designed an effective photocontrolled CTC capture/release system by combination of photochemistry and immunomagnetic separation. 7-Aminocoumarin was synthesized as the phototrigger to bridge the connection between the anti-EpCAM antibody and the magnetic beads. The coumarin moieties produced cleavage of a C-O bond under both ultraviolet (UV) and near-infrared (NIR) light illumination, breaking the bridge and releasing CTCs from the immunomagnetic beads. Compared with conventional immunomagnetic separation systems, the negative influence of absorbed immunomagnetic beads on further CTCs culture and analysis was effectively eliminated. The system can specifically recognize 10 MCF-7 cells in 1 mL of human whole blood samples with 90% efficiency and 85% purity. Under the irradiation of UV and NIR light, 73 ± 4% and 52 ± 6% of captured cells were released with a viability of 90% and 97%, respectively. Furthermore, this technique has been used to detect CTCs from whole blood of cancer patients with high purity. This study demonstrates that the photochemical-based immunomagnetic separation method for isolating, releasing and culturing CTCs from clinic patients may provide new opportunities for cancer diagnosis and personalized therapy.
稀有循环肿瘤细胞(CTC)的分离、释放和培养若能实现,可能会推动个体化抗肿瘤治疗的进展。为了在不破坏其活力的情况下实现CTC的释放,以便进一步培养和分析,我们通过光化学和免疫磁分离相结合的方式设计了一种有效的光控CTC捕获/释放系统。合成了7-氨基香豆素作为光触发剂,以桥接抗上皮细胞粘附分子(EpCAM)抗体与磁珠之间的连接。香豆素部分在紫外光(UV)和近红外光(NIR)照射下都会产生C-O键的断裂,从而打破连接并从免疫磁珠上释放出CTC。与传统的免疫磁分离系统相比,有效消除了吸附的免疫磁珠对后续CTC培养和分析的负面影响。该系统能够以90%的效率和85%的纯度特异性识别1 mL人全血样本中的10个MCF-7细胞。在紫外光和近红外光照射下,分别有73±4%和52±6%的捕获细胞被释放,活力分别为90%和97%。此外,该技术已被用于从癌症患者的全血中高纯度地检测CTC。这项研究表明,基于光化学的免疫磁分离方法用于从临床患者中分离、释放和培养CTC,可能为癌症诊断和个性化治疗提供新的机会。