bFGF expression mediated by a hypoxia-regulated adenoviral vector protects PC12 cell death induced by serum deprivation
bFGF expression mediated by a hypoxia-regulated adenoviral vector protects PC12 cell death induced by serum deprivation
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缺氧调节腺病毒载体介导的 bFGF 表达可保护血清剥夺诱导的 PC12 细胞死亡
DOI：
10.1016/j.bbrc.2009.09.077
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发表时间：
2009-12-04
影响因子：
3.1
通讯作者：
Hou, Sheng T.
中科院分区：
文献类型：
--
作者：
Hu, Hou-Wen;Li, Xiao-Kun;Hou, Sheng T.
碱性成纤维细胞生长因子（bFGF）是一种已知的神经保护剂，可对抗许多脑损伤病症，如脑缺血。然而，bFGF也调节过多的大脑发育过程，并作为一种强有丝分裂原发挥作用。因此，bFGF在脑中的不受调节的长期表达可能是潜在的致瘤性的，限制了其在脑治疗中的效用。在这里，我们报告了一个腺病毒载体（Ad-5HRE-bFGF）表达bFGF的五个低氧反应元件（5HRE）和一个最小的巨细胞病毒启动子（CMVmp）的调控下，成功的建设。在低氧室中低氧处理后，Ad-5HRE-bFGF在感染的PC12细胞中诱导bFGF蛋白和荧光标签人源化GFP（hrGFP）蛋白的显著和时间依赖性表达。与此相反，常氧处理诱发极低水平的bFGF和hrGFP表达，表明5HRE-CMVmp盒在调节响应缺氧的bFGF基因表达中是有效的。更重要的是，由Ad-5HRE-bFGF病毒载体表达的bFGF在缺氧调节下对血清剥夺引起的PC12细胞死亡具有显著的神经保护作用。总之，这些研究证明了以低氧调节的方式表达bFGF以提供神经保护的可行性。Ad-5HRE-bFGF可以进一步发展为一种有效的工具，以提供对缺氧诱导的脑疾病，如脑缺血的神经保护。皇冠版权所有（C）2009由爱思唯尔公司出版。All rights reserved.
Basic fibroblast growth factor (bFGF) is a known neuroprotectant against a number of brain injury conditions such as cerebral ischemia. However, bFGF also regulates a plethora of brain developmental processes and functions as a strong mitogen. Therefore, unregulated long-term expression of bFGF in brain may potentially be tumorigenic, limiting its utility in brain therapy. Here, we report the successful construction of an adenoviral vector (Ad-5HRE-bFGF) expressing bFGF under the regulation of five hypoxia-responsive elements (5HRE) and a minimal cytomegalovirus promoter (CMVmp). Following hypoxia treatment in a hypoxic chamber with less than 1% of oxygen, Ad-5HRE-bFGF induced a significant and time-dependent expression of bFGF protein and the fluorescent tag, humanized GFP (hrGFP) protein, in infected PC12 cells. In contrast, normoxia treatment evoked extremely low level of bFGF and hrGFP expression, demonstrating that the 5HRE-CMVmp cassette was effective in regulating the expression of bFGF gene in response to hypoxia. More importantly, bFGF expressed by the Ad-5HRE-bFGF viral vector under the regulation of hypoxia was significantly neuroprotective against PC12 cell death evoked by serum deprivation. Taken together, these studies demonstrated the feasibility to express bFGF in a hypoxia-regulated fashion to provide neuroprotection. The Ad-5HRE-bFGF can be further developed as an effective tool to provide neuroprotection against hypoxia-induced brain diseases, such as cerebral ischemia. Crown Copyright (C) 2009 Published by Elsevier Inc. All rights reserved.