Induction of vascular endothelial growth factor by nitric oxide in human glioblastoma and hepatocellular carcinoma cells
Induction of vascular endothelial growth factor by nitric oxide in human glioblastoma and hepatocellular carcinoma cells
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DOI：
10.1038/sj.onc.1201201
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发表时间：
1997-07-24
期刊：
影响因子：
8
通讯作者：
Esumi, H
中科院分区：
文献类型：
--
作者：
Chin, K;Kurashima, Y;Esumi, H
本文研究了一氧化氮（NO）对人A-172胶质母细胞瘤细胞和人肝癌细胞HepG 2血管内皮生长因子（VEGF）基因表达的影响。S-亚硝基-N-乙酰基-D，L-青霉胺（SNAP）可使两种细胞系VEGF mRNA水平升高，而A-172细胞VEGF mRNA水平的升高与VEGF蛋白的表达一致。SNAP在0.5 mM诱导的4.4和3.7 kb的VEGF mRNA表达的最大刺激后6小时约11和8倍的增加，分别高于对照水平。用另一种化学NO生成剂NOR 3也观察到类似的VEGF mRNA积累。为了评价SNAP对VEGF mRNA稳定性的影响，在用或不用0.5mM SNAP培养并用放线菌素D（25 μ g/ml）处理的A-172细胞中测量VEGF mRNA的半衰期。VEGF mRNA的半衰期被SNAP延长了约2.4倍，由SNAP诱导的VEGF表达被鸟苷酸环化酶抑制剂亚甲蓝（10 μ M）和LY-83583（1 μ M）以及蛋白质合成抑制剂放线菌酮（25 μ g/ml）抑制。这些结果表明，诱导VEGF基因的表达NO介导的鸟苷酸环化酶的活性，需要进行蛋白质合成。
We evaluated the effect of nitric oxide (NO) on vascular endothelial growth factor (VEGF) gene expression in human A-172 glioblastoma cells and human HepG2 hepatocellular carcinoma cells, The mRNA level of VEGF increased in response to S-Nitroso-N-acetyl-D,L-penicillamine (SNAP) in both cell lines, and increased in mRNA level well coincided with VEGF protein production in A-172 cells. SNAP at 0.5 mM induced maximal stimulation of 4.4 and 3.7 kb VEGF mRNA expression after 6 h about 11 and 8 fold increase, respectively above control level. Similar VEGF mRNA accumulation was observed also with NOR3, another chemical NO generator. To evaluate the effect of SNAP on VEGF mRNA stability, half-lives of VEGF mRNA were measured in A-172 cells cultured with or without 0.5 mM SNAP and treated with actinomycin D (25 mu g/ml). Half-life for VEGF mRNA was found to be prolonged about 2.4 fold by SNAP, VEGF expression induced by SNAP was inhibited by guanylate cyclase inhibitors, methylene blue (10 mu M) and LY-83583 (1 mu M), and by the protein synthesis inhibitor, cycloheximide (25 mu g/ml). These results suggest that induction of VEGF gene expression by NO is mediated through guanylate cyclase activity and requires on-going protein synthesis.