Hypoxia inducible factor 1-alpha regulates of platelet derived growth factor-B in human glioblastoma cells.
Hypoxia inducible factor 1-alpha regulates of platelet derived growth factor-B in human glioblastoma cells.
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发表时间：
2006
影响因子：
3.9
通讯作者：
D. Yoshida;Kyongsong Kim;M. Noha;A. Teramoto
中科院分区：
文献类型：
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作者：
D. Yoshida;Kyongsong Kim;M. Noha;A. Teramoto
缺氧诱导因子（HIF）是一种转录因子，调节与响应缺氧应激时氧稳态相关的多个基因的表达。尽管HIF1-α和血小板衍生生长因子-B（PDGF-B）在胶质瘤组织中表达，并且与介导血管内皮生长因子（VEGF）活性的肿瘤血管生成密切相关，但它们的直接关系尚未阐明。本研究的目的是探讨 HIF1-α 是否调节 PDGF-B 表达。将人胶质母细胞瘤细胞系 U87MG、U251MG 和 A172 暴露于 1-21% 的氧气中 24 小时。采用实时RT-PCR定量分析PDGF-B mRNA表达量，通过流式细胞仪支持的计算机图像分析测定细胞内蛋白水平，检测细胞内PDGF-B，并通过ELIA测定分泌的PDGF-B蛋白浓度。我们还在用靶向 HIF1-α mRNA 的短干扰 RNA (siRNA) 转染细胞后进行了测定。相对 PDGF-B mRNA 和 PDGF-B 蛋白的分泌在 1% 氧气下显着升高。在 1% 氧气下转染 HIF1-α siRNA 后，PDGF-B 表达在 mRNA 水平上显着受到抑制。我们的研究结果表明，HIF1-α 上调人胶质母细胞瘤细胞中 PDGF-B 的表达，并表明 siRNA 技术在胶质母细胞瘤细胞系中的可行性。
Hypoxia inducible factors (HIF) are transcription factors regulating expression of several genes related to oxygen homeostasis in response to hypoxic stress. Although HIF1-alpha and platelet derived growth factor-B (PDGF-B) are expressed in glioma tissue and closely related to tumor angiogenesis mediating vascular endothelial growth factor (VEGF) activity, their direct relationship has not yet been clarified. The aim of this study is to investigate whether HIF1-alpha regulates PDGF-B expression. The human glioblastoma cell lines, U87MG, U251MG, and A172, were exposed to 1-21% oxygen for 24 h. PDGF-B mRNA expression were quantitatively analyzed by real time RT-PCR, their intracellular protein levels were determined by computerized image analysis supported by flow cytometry to detect intracellular PDGF-B, and the concentration of secreted PDGF-B protein was assayed by ELIA. We also assayed following transfection of the cells with short interference RNA (siRNA) targeting HIF1-alpha mRNA. Relative PDGF-B mRNA and secretion of PDGF-B protein were significantly elevated at 1% oxygen. Following transfection of HIF1-alpha siRNA at 1% oxygen, PDGF-B expression was significantly suppressed at mRNA level. Our findings indicated that HIF1-alpha up-regulated expression of PDGF-B in human glioblastoma cells and showed the feasibility of siRNA technology in glioblastoma cell lines.