Enhancement of angiogenesis through stabilization of hypoxia-inducible factor-1 by silencing prolyl hydroxylase domain-2 gene.
Enhancement of angiogenesis through stabilization of hypoxia-inducible factor-1 by silencing prolyl hydroxylase domain-2 gene.
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DOI：
10.1038/mt.2008.90
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发表时间：
2008-07
期刊：
影响因子：
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通讯作者：
Shourong Wu;N. Nishiyama;M. Kano;Y. Morishita;K. Miyazono;K. Itaka;U. Chung;K. Kataoka
中科院分区：
文献类型：
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作者：
Shourong Wu;N. Nishiyama;M. Kano;Y. Morishita;K. Miyazono;K. Itaka;U. Chung;K. Kataoka
低氧诱导因子-1(HIF-1)通过激活血管内皮生长因子(VEGF)等血管生成因子，在细胞对低氧的反应中发挥重要作用。Pro羟基酶结构域2(PHD2)蛋白通过羟化特定的Pro残基来诱导HIF-1的降解。因此，用RNA干扰技术沉默PHD2基因可能通过稳定HIF-1α来增加血管生成生长因子的表达，从而促进新生血管的形成。在这项研究中，我们已经证明了PHD2的特异性沉默足以稳定缺氧诱导因子-1的α并增加其转录活性，导致血管生成因子包括血管内皮生长因子和成纤维细胞生长因子-2的表达增加。在体外培养的人脐静脉内皮细胞中，PHD2-siRNA载体转染的NIH3T3细胞的条件培养液可促进细胞增殖。此外，将Matrigel Plug与PHD2-siRNA载体转染的NIH3T3细胞混合后，在小鼠体内观察到了活体血管生成。这些结果表明，pHD2沉默通过稳定HIF-1α诱导多种血管生成生长因子的表达，并且phd2-siRNA载体在体内的植入足以促进血管生成。根据这些发现，RNAi沉默PHD2可能为血管生成治疗提供了一种潜在的工具。
Hypoxia-inducible factor-1 (HIF-1) plays a central role in cellular response to hypoxia by activating vascular endothelial growth factor (VEGF) and other angiogenic factors. Prolyl hydroxylase domain-2 (PHD2) protein induces the degradation of HIF-1 by hydroxylating specific prolyl residues. Therefore gene silencing of PHD2 by RNA interference (RNAi) might increase the expression of angiogenic growth factors and, consequently, neoangiogenesis through the stabilization of HIF-1α. In this study we have shown that the specific silencing of PHD2 is sufficient for stabilizing HIF-1α and increasing its transcriptional activity, resulting in the increased expression of angiogenic factors including VEGF and fibroblast growth factor-2 (FGF2). Moreover, when PHD2-siRNA vector was used, the increase in VEGF secretion was observed for as long as 18 days after transfection.In vitrotreatment of human umbilical vein endothelical cells with conditioned medium from PHD2-siRNA vector-transfected NIH3T3 cells was shown to increase cell proliferation. Also,in vivoangiogenesis was observed in mice implanted with Matrigel plugs mixed with NIH3T3 cells transfected with PHD2-siRNA vector. These results indicate that PHD2 silencing induces expressions of multiple angiogenic growth factors by stabilizing HIF-1α, and that the implantation of cells transfected with PHD2-siRNA vector is sufficient to enhance angiogenesisin vivo. In the light of these findings, PHD2 silencing by RNAi might offer a potential tool for angiogenic therapy.