Zhao Wenjing, Yang Liping, Chen Xudong, Qian Hongyan, Zhang Suqing, Chen Yali, Luo Runhua, Shao Jingjing, Liu Huanliang, Chen Jianguo
Cancer Research Center Nantong, Tumor Hospital Affiliated to Nantong University, Nantong, China.
Department of Hepatobiliary Surgery, Nantong Tumor Hospital, Nantong, China.
Hepatol Res. 2018 Dec;48(13):1149-1162. doi: 10.1111/hepr.13225. Epub 2018 Aug 17.
Tumor endothelial cells (TECs) have been investigated using human tumor xenografts in mice models. In order to provide pure human TECs for the updating of clinical anti-angiogenic cancer therapy, in the present study we established a protocol of purification of TECs derived from clinical hepatocellular carcinoma (HCC) and revealed the TEC features by in vitro and in vivo assays.
We isolated TECs from fresh surgical resections of HCC by magnetic-activated cell sorting and purified by flow cytometry sorting upon CD31 expression, referred to as ECDHCCs. Next, we identified cultured ECDHCCs by morphology, phenotype, genotype, and functional assays.
The ECDHCCs appeared as Weibel-Palade bodies under electron microscopy. They expressed endothelial markers, such as CD31, CD105, and vascular endothelial growth factor receptor 2, and expressed the genes that are associated with pro-angiogenesis, especially vascular endothelial growth factor, epiregulin, and programmed cell death 10. Functionally, ECDHCCs were capable of tube formation, wound healing, and Transwell migration in vitro. These in vitro behaviors were validated by in vivo Matrigel plug assay in mice. Finally, comparison of ECDHCC with the Hep-G2 liver cancer cell line showed there was no similarity of phenotype or function between these two types of cells.
Tumor endothelial cells derived from human HCC can be isolated and purified from clinical samples by flow cytometer. They have the endothelial phenotype and morphologic features and are capable of tube formation and migration. This study provides a useful model for researchers to study tumor angiogenesis and screening of candidate targets.
利用人肿瘤异种移植小鼠模型对肿瘤内皮细胞（TECs）进行了研究。为了提供纯的人TECs以更新临床抗血管生成癌症治疗方法，在本研究中，我们建立了从临床肝细胞癌（HCC）中纯化TECs的方案，并通过体外和体内试验揭示了TECs的特征。
我们通过磁激活细胞分选从HCC新鲜手术切除组织中分离TECs，并根据CD31表达通过流式细胞术分选进行纯化，称为ECD-HCCs。接下来，我们通过形态学、表型、基因型和功能试验对培养的ECD-HCCs进行鉴定。
ECD-HCCs在电子显微镜下呈现为Weibel-Palade小体。它们表达内皮标志物，如CD31、CD105和血管内皮生长因子受体2，并表达与促血管生成相关的基因，特别是血管内皮生长因子、表皮调节素和程序性细胞死亡10。在功能上，ECD-HCCs能够在体外形成管腔、伤口愈合和Transwell迁移。这些体外行为在小鼠体内基质胶栓塞试验中得到验证。最后，将ECD-HCC与Hep-G2肝癌细胞系进行比较，结果显示这两种细胞在表型或功能上没有相似性。
人HCC来源的肿瘤内皮细胞可以通过流式细胞仪从临床样本中分离和纯化。它们具有内皮表型和形态学特征，能够形成管腔和迁移。本研究为研究人员研究肿瘤血管生成和筛选候选靶点提供了一个有用的模型。
