부산대학교 도서관

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.bbrc.2009.09.077 Byline: Hou-Wen Hu (a)(e), Xiao-Kun Li (b)(d), Rong-Yuan Zheng (c), Jian Xiao (b), Jin-Qi Zeng (c), Sheng T. Hou (b)(c)(e) Keywords: Adenovirus; bFGF; Hypoxia; Hypoxia-responsive element; Gene expression; PC12 cells; Cerebral ischemia Abbreviations: bFGF, basic fibroblast growth factor; CMVmp, a minimal cytomegalovirus promoter; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; HRE, hypoxia-responsive elements; hrGFP, humanized green florescent protein; MOI, multiplicity of infection; PI, propidium iodide Abstract: 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. Author Affiliation: (a) School of Ophthalmology and Optometry, Wenzhou Medical College, Wenzhou City, Zhejiang Province, PR China (b) Key Laboratory of Biotechnology & Pharmaceutical Engineering of Zhejiang Province, School of Pharmaceutical Science, Wenzhou Medical College, Wenzhou City, Zhejiang Province, PR China (c) Department of Neurology, The First Affiliated Hospital and Research Institute of Experimental Neurobiology, Wenzhou Medical College, Wenzhou City, Zhejiang Province, PR China (d) Engineering Research Center of Bioreactor and Pharmaceutical Development, Ministry of Education, Jilin Agricultural University, Changchun, PR China (e) Institute for Biological Sciences, National Research Council of Canada, 1200 Montreal Road, Bldg M54, Ottawa, Ont., Canada K1A 0R6 Article History: Received 17 September 2009