Burke Bernard, Giannoudis Athina, Corke Kevin P, Gill Dalvir, Wells Michael, Ziegler-Heitbrock Loems, Lewis Claire E
Tumor Targeting Group, Section of Oncology and Pathology, Division of Genomic Medicine, University of Sheffield Medical School, Sheffield, United Kingdom.
Am J Pathol. 2003 Oct;163(4):1233-43. doi: 10.1016/S0002-9440(10)63483-9.
Macrophages accumulate in ischemic areas of such pathological tissues as solid tumors, atherosclerotic plaques and arthritic joints. Studies have suggested that hypoxia alters the phenotype of macrophages in a way that promotes these lesions. However, the genes up-regulated by macrophages in such hypoxic tissues are poorly characterized. Here, we have used cDNA array hybridization to investigate the effects of hypoxia on the mRNAs of 1185 genes in primary human monocyte-derived macrophages. As shown previously in other cell types, mRNA levels for vascular endothelial growth factor (VEGF) and glucose transporter 1 (GLUT-1) were up-regulated by hypoxia. However, the mRNAs of other genes were also up-regulated including matrix metalloproteinase-7 (MMP-7), neuromedin B receptor, and the DNA-binding protein inhibitor, Id2. The promoters of GLUT-1 and MMP-7 confer hypoxic inducibility on a reporter gene in RAW 264.7 macrophages, indicating that the hypoxic up-regulation of these mRNAs may occur, at least in part, at the transcriptional level. GLUT-1 and MMP-7 mRNA were also shown to be up-regulated in hypoxic macrophages in vitro by real-time RT-PCR, and these proteins were elevated in hypoxic macrophages in vitro and in hypoxic areas of human breast tumors. The hypoxia up-regulated genes identified could be important for the survival and functioning of macrophages in hypoxic diseased tissues, and their promoters could prove useful in macrophage-delivered gene therapy.
巨噬细胞在实体瘤、动脉粥样硬化斑块和关节炎关节等病理组织的缺血区域积聚。研究表明，缺氧以促进这些病变的方式改变巨噬细胞的表型。然而，巨噬细胞在这种缺氧组织中上调的基因特征尚不明确。在这里，我们使用cDNA阵列杂交来研究缺氧对原代人单核细胞衍生巨噬细胞中1185个基因mRNA的影响。如先前在其他细胞类型中所示，缺氧会上调血管内皮生长因子（VEGF）和葡萄糖转运蛋白1（GLUT-1）的mRNA水平。然而，其他基因的mRNA也上调，包括基质金属蛋白酶-7（MMP-7）、神经降压素B受体和DNA结合蛋白抑制剂Id2。GLUT-1和MMP-7的启动子赋予RAW 264.7巨噬细胞中的报告基因缺氧诱导性，表明这些mRNA的缺氧上调可能至少部分发生在转录水平。实时RT-PCR也显示GLUT-1和MMP-7 mRNA在体外缺氧巨噬细胞中上调，并且这些蛋白在体外缺氧巨噬细胞和人乳腺肿瘤的缺氧区域中升高。所鉴定的缺氧上调基因可能对缺氧疾病组织中巨噬细胞的存活和功能很重要，并且它们的启动子可能在巨噬细胞介导的基因治疗中证明有用。
