Department of Internal Medicine V, Hematology and Oncology, Medical University of Innsbruck, 6020 Innsbruck, Austria.
Lung Cancer. 2013 Aug;81(2):252-8. doi: 10.1016/j.lungcan.2013.04.014. Epub 2013 May 10.
Genome analyses of endothelial cells identified genes specifically expressed by tumor endothelial cells, called tumor endothelial markers (TEMs). Currently there are no data available concerning the role of TEMs in non-small cell lung cancer (NSCLC). Therefore, the aim of this study was to investigate the role of TEMs in NSCLC in vitro and in vivo. First we evaluated the expression of various TEMs (Robo4, Clec14 and ECSCR) by qRT-PCR and Western blot analyses in three NSCLC cell lines (A549, Calu1, Colo699) and compared them to human umbilical vein endothelial cells (HUVECs), endothelial colony forming cells (ECFCs) and human bronchial epithelial cells (HBEpCs). Next the expression of TEMs was measured in resected tumor tissue of NSCLC patients (n = 63) by qRT-PCR and compared to adjacent non-cancerous lung tissue (n = 52). Further, immunohistochemical analysis of Robo4 expression in tumor tissue (n = 33) and adjacent non-cancerous tissue (n = 27) was performed. We found that NSCLC cell lines and HBEpC did not express TEMs on the mRNA level compared to HUVECs (p = 0.001). In the contrary, a significant up-regulation of Robo4 and Clec14 was found in tumor samples (Robo4 p = 0.03, Clec14 p = 0.002). Both facts clearly indicate that these proteins are allocated to the tumor stromal department. Correlation with clinical data showed that increased TEM expression correlated with prolonged overall survival of operated NSCLC patients (Robo4 high 120.5 vs. Robo4 low 47.6 months, Clec14 high 108.1 vs. Clec14 low 54.5 months and ECSCR high 120.5 vs. ECSCR low 42.2 months). In summary, we found that TEMs are overexpressed in NSCLC stromal tissue and that an increased TEM expression correlated with an increased overall survival in early stage NSCLC.
内皮细胞的基因组分析鉴定了肿瘤内皮细胞特有的基因，称为肿瘤内皮标记物（TEMs）。目前尚无关于 TEMs 在非小细胞肺癌（NSCLC）中的作用的数据。因此，本研究旨在研究 TEMs 在 NSCLC 中的体外和体内作用。首先，我们通过 qRT-PCR 和 Western blot 分析评估了三种 NSCLC 细胞系（A549、Calu1、Colo699）中各种 TEMs（Robo4、Clec14 和 ECSCR）的表达，并将其与人类脐静脉内皮细胞（HUVECs）、内皮集落形成细胞（ECFCs）和人支气管上皮细胞（HBEpCs）进行比较。接下来，通过 qRT-PCR 测量了 NSCLC 患者切除的肿瘤组织（n = 63）中 TEMs 的表达，并与相邻的非癌性肺组织（n = 52）进行了比较。此外，还对 33 例肿瘤组织（n = 33）和 27 例相邻非癌性组织（n = 27）中的 Robo4 表达进行了免疫组织化学分析。我们发现与 HUVECs 相比，NSCLC 细胞系和 HBEpC 在 mRNA 水平上不表达 TEMs（p = 0.001）。相反，在肿瘤样本中发现 Robo4 和 Clec14 的表达显著上调（Robo4 p = 0.03，Clec14 p = 0.002）。这两个事实清楚地表明，这些蛋白质被分配到肿瘤基质部门。与临床数据的相关性表明，TEM 表达增加与接受手术治疗的 NSCLC 患者的总生存期延长相关（Robo4 高 120.5 与 Robo4 低 47.6 个月，Clec14 高 108.1 与 Clec14 低 54.5 个月和 ECSCR 高 120.5 与 ECSCR 低 42.2 个月）。总之，我们发现 TEMs 在 NSCLC 基质组织中过度表达，并且 TEM 表达增加与早期 NSCLC 患者的总生存期延长相关。
