a nuclear coiled body component protein is involved in angiogenesis.
a nuclear coiled body component protein is involved in angiogenesis.
复制标题
核卷曲体成分蛋白参与血管生成。
DOI：
10.1111/j.1349-7006.2010.01494.x
复制
发表时间：
2010
期刊：
影响因子：
5.7
通讯作者：
M.Abe. Pigpen
中科院分区：
文献类型：
--
作者：
T.Yoshida;Y. Sato;I.Morita;M.Abe. Pigpen
我们先前报道了嘌呤霉素不敏感的亮氨酰特异性氨肽酶（PILSAP）是血管内皮生长因子（VEGF）和碱性成纤维细胞生长因子（bFGF）诱导的血管生成所必需的，并且通过PILSAP的氨肽酶活性从胚胎干细胞（ES）分化为内皮细胞。在这项研究中，我们寻找在PILSAP血管生成过程中起作用的分子。我们进行了蛋白质组分析的胚胎体（EBs）从ES细胞转染突变体PILSAP缺乏氨肽酶活性和模拟EBs的核提取物。我们鉴定了pigpen，一种67 kDa的核卷曲体组分蛋白。免疫沉淀和蛋白质印迹证明PILSAP和pigpen在内皮细胞（ECs）中的结合，并且这种相互作用被VEGF和bFGF增强。据报道，Pigpen在活跃生长的EC中表达，例如胚胎和肿瘤中的EC。然而，Pigpen是否参与血管生成尚不清楚。因此，我们通过用siRNA沉默猪圈（siPigpen）来检查猪圈对血管生成的影响。与scrambled RNA相比，siPigpen转染小鼠EC抑制增殖、迁移和网络形成。这些结果用另外两种siRNA证实。此外，在基质胶塞试验中，siPigpen抑制bFGF诱导的血管生成，将siPigpen注射到皮下植入5周龄C57/BL雄性小鼠的刘易斯肺癌细胞肿瘤中可防止肿瘤生长和肿瘤血管生成。这些数据表明，猪圈参与血管生成，猪圈可能是阻断肿瘤血管生成的靶点。（Cancer Sci2010; 101：1170-1176）
We previously reported that puromycin‐insensitive leucyl‐specific aminopeptidase (PILSAP) is required for vascular endothelial growth factor (VEGF)‐ and basic fibroblast growth factor (bFGF)‐induced angiogenesis and for endothelial differentiation from embryonic stem (ES) cells via the aminopeptidase activity of PILSAP. In this study, we searched for molecules that function during angiogenesis with PILSAP. We performed proteome analysis of nuclear extracts from embryoid bodies (EBs) made from ES cells transfected with mutant PILSAP lacking aminopeptidase activity and mock EBs. We identified pigpen, a 67‐kDa nuclear coiled body component protein. Immunoprecipitation and western blotting demonstrated the binding of PILSAP and pigpen in endothelial cells (ECs), and this interaction was enhanced by VEGF and bFGF. Pigpen was reported to be expressed in actively growing ECs such as those in embryos and tumors. However, whether Pigpen is involved in angiogenesis is not known. Therefore, we examined the effect of pigpen on angiogenesis by silencing pigpen with siRNA (siPigpen). Compared with scrambled RNA (scrPigpen), transfection of siPigpen into mouse ECs inhibited proliferation, migration, and network formation. These results were confirmed with other two siRNAs. Moreover, siPigpen suppressed bFGF‐induced angiogenesis in a Matrigel plug assay, and injection of siPigpen into Lewis lung carcinoma cell tumors implanted subcutaneously into 5‐week‐old C57/BL male mice prevented tumor growth and tumor angiogenesis. These data indicate that Pigpen is involved in angiogenesis and that pigpen may be a target for blocking tumor angiogenesis.(Cancer Sci2010; 101: 1170–1176)