The role of a transcription factor, VEZF1 in endothelial differentiation and vasculogenesis

转录因子 VEZF1 在内皮分化和血管生成中的作用

• 批准号: 13832001
• 负责人: ABE Mayumi
• 金额: $ 1.22万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13832001/
• 关键词: VEZF1; etidotheital cells; transcription factors; vaculogenesis; angiogenesis; ES cells; differentiation; VEZF; 内皮細胞; 胚性幹細胞; 血管内皮前駆細胞

1. We made VEZF1 sense and antisense expression vectors (VEZF1-S and VEZF1-AS) by inserting the VEZF1 cDNA-IRES-EGFP fragment into pHPCAG, a stable expression vector for ES cells to monitor the transfection efficiency and introduced the vector intoMG1.19, an ES cell line by a supertransfection method.2. We found that VEZF1 was expressed in MG1.19 cultured on gelatin in the presence of LIF : in the undiflerentiation system.3. Although we used IRES-EGFP in pHPCAG as a Mock at first, EGFP expression was not detected in the vector. Therefore, we use EGFP in pHPCAG as a Mock.4. EGFP was detected in Mock, VEZF1-S, and VEZF1-AS by FCM. However the expression level of VEZF1 measured by RT-PCR and Western blot was the same among these three transfectants.5. Furthermore, the expression of VEGFR2 (Flk-1), a marker of endothelial progenitor cells was the same in these three transfectants on OP9 cells without LIF: in the differentiation system.6. We found VEZF1 was essential for endothetial growth, migration, and network formation in Vitro and in Vivo (our unpublished data). However, we could not clarify the involvement of VEZF1 in endothelial differentiation as well as in angiogenesis because of failure in inhibiting VEZF1 expression by VEZF1-AS.7. We would like to try using siRNA.

1。我们通过将VEZF1 cDNA-IRES-EGFP片段插入PHPCAG中，使VEZF1有道理和反义表达载体（VEZF1-S和VEZF1-AS），这是ES细胞的稳定表达向量，以监测转染效率，并通过超级分析方法。我们发现VEZF1在LIF的存在下在明胶上培养的Mg1.19中表达：在未分类系统中。3。尽管我们首先将IRES-EGFP在PHPCAG中用作模拟，但在向量中未检测到EGFP的表达。因此，我们在PHPCAG中使用EGFP作为模拟4。 FCM在模拟，VEZF1-S和VEZF1-AS中检测到EGFP。然而，通过RT-PCR和Western印迹测得的VEZF1的表达水平在这三种转染剂中是相同的。5。此外，在没有LIF的OP9细胞上的这三种转染剂中，内皮祖细胞的表达（FLK-1）的表达相同：在分化系统中。6。我们发现VEZF1对于体外和体内（我们未发表的数据）中的内刺生长，迁移和网络形成至关重要。但是，由于VEZF1-AS.7抑制VEZF1表达的失败，我们无法阐明VEZF1参与内皮分化以及血管生成。我们想尝试使用siRNA。

项目成果

Abe M., Inoue D., Matsunaga K., Ohizumi Y., Ueda H., Asano T., Murakami M., Sato Y.: "Goniodomin A, an antifungal polyether macrolide, exhibits antiangiogenic activities via inhibiting actin reorganization of endothelial cells"J.Cell.Physiol.. 190. 109-11

Abe M.、Inoue D.、Matsunaga K.、Ohizumi Y.、Ueda H.、Asano T.、Murakami M.、Sato Y.：“Goniodomin A 是一种抗真菌聚醚大环内酯，通过抑制内皮肌动蛋白重组而表现出抗血管生成活性。

T.Nakagawa: "HEX Acts as a Negative Regulator of Angibgenesis by Modulating the Expression of Angiogenesis-Related Gene in Endothelial Cells In Vitro"Arterioscle. Thromb. Vase. Bioh.. 23. 231-237 (2003)

T.Nakakawa：“HEX 通过调节体外内皮细胞中血管生成相关基因的表达，充当血管生成的负调节剂”。

M.Oikawa: "Hypoxia induces transcription factor ETS-1 via the activity of Hypoxia-inducible factor-1"Biochem Biophys Res Commun.. 289(1). 39-43 (2001)

M.Oikawa：“缺氧通过缺氧诱导因子-1 的活性诱导转录因子 ETS-1”Biochem Biophys Res Commun.. 289(1)。

M.Abe: "Augmented binding and activation of latent transforming growth fector-β by a tryptic fragment of latency associated peptide"Endothelium. 9. 25-36 (2002)

M.Abe：“通过潜伏相关肽的胰蛋白酶片段增强潜伏转化生长因子-β的结合和激活”Endothelium。

Y.Terai: "Vascular smooth muscle cell growth-promoting factor/F-spondin inhibits angiogenesis via the blockade of integrin avβ3 on vascular endothelial cells"J. Cell. Physiol. 188. 394-402 (2001)

Y. Terai：“血管平滑肌细胞生长促进因子/F-spondin 通过阻断血管内皮细胞上的整合素 avβ3 抑制血管生成”J. Physiol。