Endothelial cells of human colorectal cancer and healthy colon reveal phenotypic differences in culture
Endothelial cells of human colorectal cancer and healthy colon reveal phenotypic differences in culture
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DOI：
10.1038/labinvest.3700671
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发表时间：
2007-11
影响因子：
5
通讯作者：
V. Schellerer;R. Croner;Kristina Weinländer;W. Hohenberger;M. Stürzl;E. Naschberger
中科院分区：
文献类型：
--
作者：
V. Schellerer;R. Croner;Kristina Weinländer;W. Hohenberger;M. Stürzl;E. Naschberger
抗血管生成药物已成功用于结直肠癌的治疗。同一患者未受累结肠组织的活肿瘤内皮细胞(TEC)和正常内皮细胞(NEC)在体外尚不能用于优化治疗策略。因此，我们的目标是建立分离TEC和NEC的方案。通过对8名患者的结直肠癌和健康结肠组织样本进行磁性细胞分选，获得了非常高纯度的这些细胞。Tec和NEC表达CD31、CD105、VE-cadherin、VCAM-1、ICAM-1和E-选择素，在基底膜提取液中形成毛细血管，并能摄取乙酰化低密度脂蛋白。泊多拉宁、CD45、CD68和细胞角蛋白-20均为阴性，提示血管内皮细胞系。Von Willebrand因子(VWF)在8种培养的NEC中有5种呈强阳性表达，而在所有体外培养的TEC中均不表达或仅有微弱表达。在组织水平上，TEC和NEC细胞内也检测到低浓度的vWF。这表明TEC和NEC在体内表现出的差异在培养中稳定地保持不变。分离的培养物可能为阐明表观遗传学对肿瘤血管生成的影响和优化抗血管生成治疗提供有用的体外模型。
Antiangiogenic drugs have been used successfully for the treatment of colorectal cancer (CRC). Viable tumor endothelial cells (TEC) and normal endothelial cells (NEC) of uninvolved colon tissue of the same patient have not been available to optimize treatment strategies in vitro. Therefore, our target was to establish a protocol for the isolation of TEC and NEC. These cells were isolated with very high purity via magnetic cell sorting of tissue samples obtained from CRC and healthy colon of eight patients. TEC and NEC expressed CD31, CD105, VE-cadherin, VCAM-1, ICAM-1 and E-selectin, formed capillaries in basal membrane extract and were able to take up acetylated low-density lipoprotein. They were negative for podoplanin, CD45, CD68 and cytokeratin-20 indicating blood vessel endothelial lineage. Intense staining of von Willebrand factor (vWF) was observed in five of eight NEC cultures, whereas vWF was absent or only slightly expressed in all TEC cultures in vitro. Low intracellular concentration of vWF was also detected in TEC and NEC at the tissue level. This demonstrated that differences exhibited by TEC and NEC in vivo are stably perpetuated in culture. The isolated cultures may provide a useful in vitro model to elucidate epigenetic effects on angiogenesis in cancer and to optimize antiangiogenic therapy.