Department of Organ Transplantation, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai, China.
Jukang (Shanghai) Biotechnology Co. Ltd., 28, Xiangle Rd., Shanghai, 201800, China.
J Nanobiotechnology. 2021 Mar 16;19(1):74. doi: 10.1186/s12951-021-00818-3.
This research was to develop a special method for enriching Circulating tumor cells (CTCs) of Hepatocellular carcinoma (HCC) by Glypican-3 immunoliposomes (GPC3-IML), and to analyze the correlation between the CTCs count and tumor malignancy, as well as to investigate the mutation characteristics of CTC-derived NGS.
In this study characterization of physical parameters was performed with the preparation of GPC3-IML. CTCs in peripheral blood of HCC patients were further separated and identified. Immunofluorescence was used to identify CTCs for further counting. By this means, the correlation between CTCs count and clinicopathological features was analyzed, and the genetic mutation characteristics of NGS derived from CTCs were investigated and compared with that of tissue NGS. Results showed that compared with EpCAM and vimentin, GPC-3 had a stronger CTCs separation ability. There was a correlation between "positive" count of CTCs (≥ 5 PV-CTC per 7.5 ml blood) and BCLC stage (P = 0.055). The result of CTC-NGS was consistent with that of tissue-NGS in 60% cases, revealing that KMT2C was a common highly-frequent mutated gene.
The combination of immunomagnetic separation of CTCs and anti-tumor marker identification technology can be regarded as a new technology of CTCs detection in peripheral blood of patients with HCC. Trial registration EHBHKY2020-k-024. Registered 17 August 2020-Retrospectively registered.
本研究旨在通过 GPC3 免疫脂质体(GPC3-IML)开发一种富集肝细胞癌(HCC)循环肿瘤细胞(CTC)的特殊方法,并分析 CTC 计数与肿瘤恶性程度的相关性,以及研究 CTC 衍生 NGS 的突变特征。
本研究通过制备 GPC3-IML 对其物理参数进行了表征。进一步分离和鉴定 HCC 患者外周血中的 CTC。通过免疫荧光鉴定 CTC 并进一步计数。通过这种方法,分析了 CTC 计数与临床病理特征的相关性,并研究和比较了 CTC 衍生 NGS 的遗传突变特征与组织 NGS。结果表明,与 EpCAM 和波形蛋白相比,GPC-3 具有更强的 CTC 分离能力。“阳性”CTC 计数(每 7.5ml 血液中≥5 PV-CTC)与 BCLC 分期之间存在相关性(P=0.055)。CTC-NGS 的结果在 60%的病例中与组织-NGS 一致,表明 KMT2C 是一种常见的高频突变基因。
CTC 免疫磁分离与肿瘤标志物识别技术相结合,可作为一种检测 HCC 患者外周血中 CTC 的新技术。试验注册号 EHBHKY2020-k-024。于 2020 年 8 月 17 日注册-回顾性注册。