Anti-angiogenic action of the C-terminal domain of tenomodulin that shares homology with chondromodulin-I
Anti-angiogenic action of the C-terminal domain of tenomodulin that shares homology with chondromodulin-I
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DOI：
10.1242/jcs.01112
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发表时间：
2004-06-01
影响因子：
4
通讯作者：
Shukunami, C
中科院分区：
文献类型：
--
作者：
Oshima, Y;Sato, K;Shukunami, C
Tenomodulin（TeM）是一种II型跨膜糖蛋白，其含有与软骨源性血管生成抑制剂软骨调节素-I（ChM-I）的成熟分泌形式同源的C-末端结构域。TeM转录本已在少血管组织如肌腱和韧带中发现，但TeM的生物活性尚未完全研究。使用腺病毒表达系统，我们利用人TeM C-末端116个氨基酸（Ad-shTeM）在人脐静脉内皮细胞（HUVECs）中的强制表达和随后的分泌来评估TeM的抗血管生成特性。作为比较研究，人ChM-I前体（Ad-shChM-I）的C-末端120个氨基酸在HUVEC中类似地表达。当在基质胶中培养时，Ad-shTeM和Ad-shChM-I的转导导致HUVECs的管形成活性的显著损害。类似地，与来自用对照载体转染的COS 7细胞或来自未转染细胞的培养基相比，来自用编码shTeM或shChM-I的质粒DNA转染的COS 7细胞的条件培养基抑制HUVEC的管形成。在用Ad-shTeM或Ad-shChM-I感染HUVECs后，由血管内皮生长因子（VEGF）刺激的DNA合成降低至正常水平的40-50%。此外，在改良的Boyden室测定中，HUVEC响应VEGF的迁移在Ad-shTeM或Ad-shChM-I转导后受到显著影响，并且发现这些转导的HUVEC在I型胶原或纤连蛋白上良好地扩散，但在玻连蛋白上不扩散。此外，Ad-shTeM或Ad-shChM-I在人黑素瘤细胞中的转导导致与体内血管密度降低相关的肿瘤生长抑制。因此，我们已经证明，类似于ChM-I，TeM的C-末端结构域在以分泌形式表达时表现出抗血管生成和抗肿瘤活性。
Tenomodulin (TeM) is a type II transmembrane glycoprotein that contains a C-terminal domain with homology to the mature, secreted form of chondromodulin-I (ChM-I), a cartilage-derived angiogenesis inhibitor. TeM transcripts have been found in hypovascular tissues such as tendons and ligaments but the biological activity of TeM has not yet been fully explored. Using an adenovirus expression system, we utilized the forced expression and subsequent secretion of the human TeM C-terminal 116 amino acids (Ad-shTeM) in human umbilical vein endothelial cells (HUVECs) to assess the anti-angiogenic properties of TeM. The C-terminal 120 amino acids of the human ChM-I precursor (Ad-shChM-I) was similarly expressed in HUVECs as a comparison study. Transduction of both Ad-shTeM and Ad-shChM-I resulted in significant impairment of the tube-forming activity of HUVECs, when cultured in Matrigel. Similarly, conditioned medium from COS7 cells, transfected with plasmid DNA encoding shTeM or shChM-I, inhibited tube formation of HUVECs when compared to medium derived from either COS7 cells transfected with control vector or from non-transfected cells. Upon infection of HUVECs with Ad-shTeM or Ad-shChM-I, DNA synthesis stimulated by vascular endothelial growth factor (VEGF) was reduced to 40-50% of normal levels. Additionally, in a modified Boyden chamber assay, migration of HUVECs in response to VEGF was significantly affected following transduction of either Ad-shTeM or Ad-shChM-I and these transduced HUVECs were found to spread well on type I collagen or fibronectin, but not on vitronectin. Furthermore, the transduction of either Ad-shTeM or Ad-shChM-I in human melanoma cells resulted in suppression of tumor growth in association with decreased vessel density in vivo. Hence, we have demonstrated that, similarly to ChM-I, the C-terminal domain of TeM exhibits both anti-angiogenic and anti-tumor activities when expressed in a secreted form.