Lei Yongrong, Wang Xishu, Sun Heng, Fu Yuna, Tian Yichen, Yang Ludi, Wang Jianhua, Xia Feng
Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.
Key Laboratory of Hepatobiliary and Pancreatic Surgery, Institute of Hepatobiliary Surgery, Southwest Hospital, the First Hospital Affiliated to AMU (Southwest Hospital), Chongqing, China.
Front Oncol. 2021 May 27;11:601668. doi: 10.3389/fonc.2021.601668. eCollection 2021.
Cancer stem cells (CSCs) and Circulating tumor cells (CTCs) have been proposed as fundamental causes for the recurrence of hepatocellular carcinoma (HCC). CTCs isolated from patients with HCC illustrate a unique Nanog expression profile analysis. The aim of this study was to enhance the prediction of recurrence and prognosis of the CTC phenotype in patients with HCC by combining Nanog expression into a combined forecasting model.
We collected 320 blood samples from 160 patients with HCC cancer before surgery and used CanPatrol™ CTC enrichment technology and in situ hybridization (ISH) to enrich and detect CTCs and CSCs. Nanog expression in all CTCs was also determined. In addition, RT-PCR and immunohistochemistry were used to study the expression of Nanog, E-Cadherin, and N-Cadherin in liver cancer tissues and to conduct clinical correlation studies.
The numbers of CTCs and CTCs were strongly correlated with postoperative HCC recurrence (CTC number (P = 0.03), the total number of mixed CTCS (P = 0.02), and Nanog> 6.7 (P = 0.001), with Nanog > 6.7 (P = 0.0003, HR = 2.33) being the most crucial marker. There are significant differences in the expression of Nanog on different types of CTC: most Epithelial CTCs do not express Nanog, while most of Mixed CTC and Mesenchymal CTC express Nanog, and their positive rates are 38.7%, 66.7%, and 88.7%, respectively, (P=0.0001). Moreover, both CTC (≤/> 13.3) and Nanog (≤/>6.7) expression were significantly correlated with BCLC stage, vascular invasion, tumor size, and Hbv-DNA (all P < 0.05). In the young group and the old group, patients with higher Nanog expression had a higher recurrence rate. (P < 0.001).
The number of Nanog-positive cells showed positive correlation with the poor prognosis of HCC patients. The detection and analysis of CTC markers (EpCAM and CK8, 18, CD45 Vimentin,Twist and 19) and CSCs markers (NANOG) are of great value in the evaluation of tumor progression.
癌症干细胞（CSCs）和循环肿瘤细胞（CTCs）被认为是肝细胞癌（HCC）复发的根本原因。从HCC患者中分离出的CTCs显示出独特的Nanog表达谱分析。本研究的目的是通过将Nanog表达纳入联合预测模型，提高对HCC患者CTCs表型复发和预后的预测。
研究对象、材料与方法：我们收集了160例HCC患者术前的320份血样，采用CanPatrol™ CTC富集技术和原位杂交（ISH）对CTCs和CSCs进行富集和检测。同时测定所有CTCs中Nanog的表达。此外，采用RT-PCR和免疫组化方法研究肝癌组织中Nanog、E-钙黏蛋白和N-钙黏蛋白的表达，并进行临床相关性研究。
CTCs数量和混合CTCs总数与HCC术后复发密切相关（CTCs数量（P = 0.03），混合CTCs总数（P = 0.02），以及Nanog> 6.7（P = 0.001），其中Nanog > 6.7（P = 0.0003，HR = 2.33）是最关键的标志物。不同类型CTCs上Nanog的表达存在显著差异：大多数上皮性CTCs不表达Nanog，而大多数混合性CTCs和间充质性CTCs表达Nanog，其阳性率分别为38.7%、66.7%和88.7%，（P = 0.0001）。此外，CTCs（≤/> 13.3）和Nanog（≤/>6.7）表达均与BCLC分期、血管侵犯、肿瘤大小和乙肝病毒DNA显著相关（均P < 0.05）。在年轻组和老年组中，Nanog表达较高的患者复发率较高。（P < 0.001）。
Nanog阳性细胞数量与HCC患者预后不良呈正相关。CTCs标志物（EpCAM和CK8、18、CD45、波形蛋白、Twist和19）和CSCs标志物（NANOG）的检测和分析在评估肿瘤进展方面具有重要价值。
