Expression patterns of the hypoxia-related genes osteopontin, CA9, erythropoietin, VEGF and HIF-1α in human glioma in vitro and in vivo
Expression patterns of the hypoxia-related genes osteopontin, CA9, erythropoietin, VEGF and HIF-1α in human glioma in vitro and in vivo
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DOI：
10.1016/j.radonc.2007.05.003
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发表时间：
2007-06-01
影响因子：
5.7
通讯作者：
Vordermark, Dirk
中科院分区：
文献类型：
--
作者：
Said, Harun M.;Hagemann, Carsten;Vordermark, Dirk
背景与目的：为了确定胶质母细胞瘤（GBM）肿瘤缺氧的分子标记和潜在的治疗靶点，我们在体外研究了人GBM细胞系和体内肿瘤样品中骨桥蛋白（OPN）、碳酐酶9 （CA9）、促红细胞生成素（EPO）、血管内皮生长因子（VEGF）和缺氧诱导因子-1 α (HIF-1a)的缺氧相关表达，并与低级别星形细胞瘤（LGA）进行了比较。材料与方法：采用半定量RT-PCR方法，分析体外缺氧条件下（5%、1%或0.1% 02条件下1、6或24 h） LGA和GBM两组患者（n = 15）肿瘤样本中gmg、U373和U251细胞中缺氧诱导基因OPN、CA9、EPO、VEGF和HIF-1a的表达情况。选定条件和代表性肿瘤样本也通过Western blot在蛋白水平上进行评价。结果：OPN和CA9 mRNA的上调与体外缺氧的严重程度和持续时间有关。在肿瘤样本中，OPN的平均表达水平（LGA vs. GBM，归一化为正常脑中的平均表达）为1.71 vs. 4.57 （p < 0.001）。CA9为11比3.35 (p < 0.001)， Epo为2.79比5.28（无统计学意义），1。VEGF为13比2.0 (p = 0.007)， HIF-1a为0.97比0.97 （n.s s）。在肿瘤样本中，GBM显示出特别强的OPN蛋白表达。结论：在一组已知的缺氧诱导基因中，OPN和CA9在人GBM细胞系中被体外缺氧诱导的一致性最强，在患者GBM肿瘤组织中表达的特异性最强，这使得这两个基因成为缺氧指导治疗方法的有吸引力的靶点。2007爱思唯尔爱尔兰有限公司版权所有。
Background and purpose: To identify molecular markers of tumor hypoxia and potential therapeutic targets in glioblastorna (GBM), we investigated the hypoxia-related expression of osteopontin (OPN), carbonic anhydrase 9 (CA9), erythropoietin (EPO), vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1 alpha (HIF-1a) in vitro in human GBM cell lines and in vivo in human tumor samples of GBM, compared to low-grade astrocytoma (LGA).Materials and methods: Expression of the hypoxia-induced genes OPN, CA9, EPO, VEGF and HIF-1a was analyzed in three GBM cell lines, GaMG, U373 and U251, under in vitro hypoxia (1, 6 or 24 h at 5%, 1% or 0.1% 02) and in tumor samples from two patient groups with LGA and GBM (n = 15 each), at the mRNA level (semiquantitative RT-PCR). Selected conditions and representative tumor samples were also evaluated at the protein level by Western blot.Results: OPN and CA9 mRNA was most consistently upregulated in relation to severity and duration of in-vitro hypoxia. In tumor samples, mean expression levels (LGA vs. GBM, normalized to mean expression in normal brain) were 1.71 vs. 4.57 (p < 0.001) for OPN, 1. 11 vs. 3.35 (p < 0.001)for CA9, 2.79 vs. 5.28 (not significant, n.s.)for Epo, 1. 13 vs. 2.0 (p = 0.007) for VEGF and 0.97 vs. 0.97 (n.s.) for HIF-1a. In tumor samples, GBM showed a particularly strong protein expression of OPN.Conclusions: Among a panel of known hypoxia-inducible genes, OPN and CA9 emerge as most consistently induced by in vitro hypoxia in human GBM cell lines and most specifically expressed in patient GBM tumor tissue, rendering these two genes attractive targets for hypoxia-directed treatment approaches. (C) 2007 Elsevier Ireland Ltd. All rights reserved.