Development of Immortalized Human Tumor Endothelial Cells from Renal Cancer
Development of Immortalized Human Tumor Endothelial Cells from Renal Cancer
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DOI：
10.3390/ijms20184595
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发表时间：
2019-09-02
影响因子：
5.6
通讯作者：
Hida, Kyoko
中科院分区：
文献类型：
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作者：
Maishi, Nako;Kikuchi, Hiroshi;Hida, Kyoko
肿瘤血管生成研究和抗血管生成药物的开发利用了培养的内皮细胞，包括人的微血管内皮细胞等。然而，已有研究报道肿瘤内皮细胞(TECs)不同于正常内皮细胞(NECs)。为了从功能上验证抗血管生成药物，培养的TECs是不可或缺的工具，但还没有商业化。原代人TEC只能从手术标本中获得少量的细胞，由于其细胞衰老，在体外寿命较短。我们通过感染产生猴病毒40大T抗原的慢病毒和人端粒酶逆转录酶，建立了永生化的人TECs(h-imTECs)及其正常对应物(h-imNECs)，以克服复制障碍。这些内皮细胞表现出延长的寿命，并保留了其特有的内皮细胞形态、内皮细胞标志物的表达和管状形成能力。此外，h-imTECs还表现出其特有的TEC特征，如增殖增加、TEC标志物上调等。贝伐单抗，一种抗血管生成药物，显著降低了h-Imtec存活率，而同样的治疗未能改变永生化的NEC存活率。因此，这些h-imTECs可能是一种有价值的药物筛选工具，可用于开发针对TECs的新型治疗药物或在肿瘤血管生成研究中进行功能生物学检测。
Tumor angiogenesis research and antiangiogenic drug development make use of cultured endothelial cells (ECs) including the human microvascular ECs among others. However, it has been reported that tumor ECs (TECs) are different from normal ECs (NECs). To functionally validate antiangiogenic drugs, cultured TECs are indispensable tools, but are not commercially available. Primary human TECs are available only in small quantities from surgical specimens and have a short life span in vitro due to their cellular senescence. We established immortalized human TECs (h-imTECs) and their normal counterparts (h-imNECs) by infection with lentivirus producing simian virus 40 large T antigen and human telomerase reverse transcriptase to overcome the replication barriers. These ECs exhibited an extended life span and retained their characteristic endothelial morphology, expression of endothelial marker, and ability of tube formation. Furthermore, h-imTECs showed their specific characteristics as TECs, such as increased proliferation and upregulation of TEC markers. Treatment with bevacizumab, an antiangiogenic drug, dramatically decreased h-imTEC survival, whereas the same treatment failed to alter immortalized NEC survival. Hence, these h-imTECs could be a valuable tool for drug screening to develop novel therapeutic agents specific to TECs or functional biological assays in tumor angiogenesis research.