Department of Pharmaceutics, Key Laboratory of Chemical Biology (Ministry of Education), School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, 250012, People's Republic of China.
Department of Neurosurgery, Shandong Provincial Hospital Affiliated with Shandong First Medical University, Jinan, 250021, People's Republic of China.
Int J Nanomedicine. 2021 Jun 18;16:4161-4173. doi: 10.2147/IJN.S307691. eCollection 2021.
Specific targeting receptors for efficiently capturing and applicable nanodevice for separating and instant observing of circulating tumour cells (CTC) are critical for early diagnosis of cancer. However, the existing CTC detection system based on epithelial cell adhesion molecule (EpCAM) was seriously limited by low expression and poor specificity of targeting receptors, and not instant observation in clinical application.
Herein, an alternative glypican-3 (GPC3)-based immunomagnetic fluorescent system (C6/MMSN-GPC3) for high-specific isolation and instant observation of CTC from hepatocellular carcinoma (HCC) patients' peripheral blood was developed. The high-specific HCC targeting receptor, GPC3, was employed for improving the sensitivity and accuracy in CTC detection. GPC3 monoclonal antibody (mAb) was linked to immunomagnetic mesoporous silica for specific targeting capture and separate CTC, and fluorescent molecule coumarin-6 (C6) was loaded for instant detection of CTC.
The cell recovery (%) of C6/MMSN-GPC3 increased in 10 HL-60 cells (from 49.7% to 83.0%) and in whole blood (from 42% to 80.3%) compared with MACS Beads. In clinical samples, the C6/MMSN-GPC3 could capture more CTC in the 13 cases of HCC patients and the capture efficiency was improved by 83.3%-350%. Meanwhile, the capture process of C6/MMSN-GPC3 was harmless, facilitating for the subsequent culture. Significantly, the C6/MMSN-GPC3 achieved the high-specific isolation and instant observation of CTC from HCC patients' blood samples, and successfully separated CTC from one patient with early stage of HCC (Stage I) and one post-surgery patient, further indicating the potential ability of C6/MMSN-GPC3 for HCC early diagnosis and prognosis evaluation.
Our study provides a feasible glypican-3 (GPC3)-based immunomagnetic fluorescent system (C6/MMSN-GPC3) for high-specific isolation and instant observation of HCC CTC.
高效捕获和适用的纳米器件对于分离和即时观察循环肿瘤细胞（CTC）的特异性靶向受体是癌症早期诊断的关键。然而，现有的基于上皮细胞黏附分子（EpCAM）的 CTC 检测系统受到靶向受体表达率低和特异性差的严重限制，并且在临床应用中无法即时观察。
本文开发了一种基于磷脂酰聚糖-3（GPC3）的免疫磁荧光系统（C6/MMSN-GPC3），用于从肝癌（HCC）患者外周血中高特异性分离和即时观察 CTC。使用高特异性 HCC 靶向受体 GPC3 提高 CTC 检测的灵敏度和准确性。将 GPC3 单克隆抗体（mAb）连接到免疫磁介孔硅上，用于特异性靶向捕获和分离 CTC，并装载荧光分子香豆素-6（C6）用于即时检测 CTC。
与 MACS 磁珠相比，C6/MMSN-GPC3 可使 10 个 HL-60 细胞（从 49.7%增加到 83.0%）和全血（从 42%增加到 80.3%）的细胞回收率（%）增加。在临床样本中，C6/MMSN-GPC3 可以在 13 例 HCC 患者中捕获更多的 CTC，并且捕获效率提高了 83.3%-350%。同时，C6/MMSN-GPC3 的捕获过程是无害的，便于后续培养。值得注意的是，C6/MMSN-GPC3 能够从 HCC 患者的血液样本中高特异性分离和即时观察 CTC，并成功从一名早期 HCC（I 期）和一名手术后患者中分离出 CTC，进一步表明 C6/MMSN-GPC3 具有用于 HCC 早期诊断和预后评估的潜力。
本研究提供了一种可行的基于磷脂酰聚糖-3（GPC3）的免疫磁荧光系统（C6/MMSN-GPC3），用于高特异性分离和即时观察 HCC CTC。
