Hypoxia-regulated neurotrophin-3 expression by multicopy hypoxia response elements reduces apoptosis in PC12 cells

Zhang,Junfeng; Shi,Qindong; Chen,Xinlin; Yang,Pengbo; Qi,Cunfang; Zhang,Jianshui; Lu,Haixia; Liu,Jianxin; Jiao,Qian; Zhao,Lingyu; Zhao,Bingqiao; Zheng,Ping; Liu,Yong

doi:10.3892/ijmm.2012.1119

Hypoxia-regulated neurotrophin-3 expression by multicopy hypoxia response elements reduces apoptosis in PC12 cells

Authors:
- Junfeng Zhang
- Qindong Shi
- Xinlin Chen
- Pengbo Yang
- Cunfang Qi
- Jianshui Zhang
- Haixia Lu
- Jianxin Liu
- Qian Jiao
- Lingyu Zhao
- Bingqiao Zhao
- Ping Zheng
- Yong Liu
View Affiliations

Affiliations: Institute of Neurobiology, Environment and Genes Related to Diseases Key Laboratory of Education Ministry, Xi'an Jiaotong University College of Medicine, Xi'an, Shaanxi 710061, P.R. China, The State Key Laboratory of Medical Neurobiology, Shanghai 200032, P.R. China
Published online on: September 6, 2012 https://doi.org/10.3892/ijmm.2012.1119
Pages: 1173-1179

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

We have previously reported that 5 copies of the hypoxia response element (HRE) can conditionally regulate brain-derived neurotrophic factor gene expression under hypoxic/ischemic conditions in mice. In the present study, we investigated the controlled expression of neurotrophin-3 (NT-3) by HRE under hypoxic conditions and determined the protective effects of conditionally expressed NT-3 on hypoxia-induced apoptosis in PC12 cells. Five copies of the HRE (5HRE) and the simian virus 40 minimal promoter (SV40mp) were employed to construct a cassette, and transfer of therapeutic gene, NT-3, into PC12 cells was achieved using a retroviral vector. Our results showed that the retroviral vector, pLNC-5HRE-NT3, was successfully constructed and transfected into PC12 cells. Compared with normal conditions, in which NT-3 was expressed at low levels, the expression of NT-3 significantly increased under hypoxic conditions in 5HRE-NT3 transgenic PC12 cells (P<0.05). By contrast, in NT-3 transgenic PC12 cells without HRE, we found no significant difference in NT-3 expression between the normoxic and hypoxic groups. The conditional adjustment of NT-3 expression by 5HRE significantly reduced apoptosis induced by hypoxia in 5HRE-NT3 transgenic PC12 cells (P<0.05) but not in 5HRE-enhanced green fluorescent protein (EGFP) transgenic PC12 cells and PC12 cells without gene transfer. In addition, the hypoxia-induced upregulation of both p38 and caspase-3 activities was suppressed in 5HRE-NT3 transgenic PC12 cells under hypoxic conditions (P<0.05). Taken together, these results demonstrate that 5HRE-SV40mp regulates NT-3 gene expression in response to hypoxia in PC12 cells. The data presented in this study may prove useful in future gene therapy studies for the treatment of ischemic diseases.

View Figures

View References

1.	A BeenkenM MohammadiThe FGF family: biology, pathophysiology and therapyNat Rev Drug Discov8235253200910.1038/nrd279219247306
2.	RJ LeeML SpringerWE Blanco-BoseR ShawPC UrsellHM BlauVEGF gene delivery to myocardium: deleterious effects of unregulated expressionCirculation102898901200010.1161/01.CIR.102.8.89810952959
3.	Q ShiP ZhangJ ZhangAdenovirus-mediated brain-derived neurotrophic factor expression regulated by hypoxia response element protects brain from injury of transient middle cerebral artery occlusion in miceNeurosci Lett465220225200910.1016/j.neulet.2009.08.049
4.	F ShenH SuY FanAdeno-associated viral-vector-mediated hypoxia-inducible vascular endothelial growth factor gene expression attenuates ischemic brain injury after focal cerebral ischemia in miceStroke3726012606200610.1161/01.STR.0000240407.14765.e8
5.	T ShibataAJ GiacciaJM BrownDevelopment of a hypoxia-responsive vector for tumor-specific gene therapyGene Ther7493498200010.1038/sj.gt.330112410757022
6.	H SuJ Arakawa-HoytYW KanAdeno-associated viral vector-mediated hypoxia response element-regulated gene expression in mouse ischemic heart modelProc Natl Acad Sci USA9994809485200210.1073/pnas.13227529912084814
7.	GL SemenzaTargeting HIF-1 for cancer therapyNat Rev Cancer3721732200310.1038/nrc1187
8.	NV IyerLE KotchF AganiCellular and developmental control of O₂ homeostasis by hypoxia-inducible factor 1 alphaGenes Dev1214916219989436976
9.	A PatapoutianLF ReichardtTrk receptors: mediators of neurotrophin actionCurr Opin Neurobiol11272280200110.1016/S0959-4388(00)00208-711399424
10.	X LiZ YangA ZhangThe effect of neurotrophin-3/chitosan carriers on the proliferation and differentiation of neural stem cellsBiomaterials3049784985200910.1016/j.biomaterials.2009.05.04719539985
11.	H LuM LiT SongRetrovirus delivered neurotrophin-3 promotes survival, proliferation and neuronal differentiation of human fetal neural stem cells in vitroBrain Res Bull77158164200810.1016/j.brainresbull.2008.02.03719875351
12.	HX LuZM HaoQ JiaoNeurotrophin-3 gene transduction of mouse neural stem cells promotes proliferation and neuronal differentiation in organotypic hippocampal slice culturesMed Sci Monit17BR305BR311201122037732
13.	ZH ZhangRZ WangGL LiTransplantation of neural stem cells modified by human neurotrophin-3 promotes functional recovery after transient focal cerebral ischemia in ratsNeurosci Lett444227230200810.1016/j.neulet.2008.08.049
14.	B CristofaroOA StoneA CaporaliNeurotrophin-3 is a novel angiogenic factor capable of therapeutic neovascularization in a mouse model of limb ischemiaArterioscler Thromb Vasc Biol3011431150201010.1161/ATVBAHA.109.20546820360537
15.	K ShimazuM ZhaoK SakataNT-3 facilitates hippo-campal plasticity and learning and memory by regulating neurogenesisLearn Mem13307315200610.1101/lm.7600616705139
16.	T ShibataAJ GiacciaJM BrownHypoxia-inducible regulation of a prodrug-activating enzyme for tumor-specific gene therapyNeoplasia44048200210.1038/sj.neo.790018911922390
17.	H RuanH SuL HuKR LambornYW KanDF DeenA hypoxia-regulated adeno-associated virus vector for cancer-specific gene therapyNeoplasia3255263200110.1038/sj.neo.790015711494119
18.	N MoriR SteinO SigmundDJ AndersonA cell type-preferred silencer element that controls the neural-specific expression of the SCG10 geneNeuron4583594199010.1016/0896-6273(90)90116-W2322462
19.	D HuangA DesboisST HouA novel adenoviral vector which mediates hypoxia-inducible gene expression selectively in neuronsGene Ther1213691376200510.1038/sj.gt.3302538
20.	HW HuXK LiRY ZhengJ XiaoJQ ZengST HoubFGF expression mediated by a hypoxia-regulated adenoviral vector protects PC12 cell death induced by serum deprivationBiochem Biophys Res Commun390115120200910.1016/j.bbrc.2009.09.07719782044
21.	JA HillionK TakahashiD MaricC RuetzlerJL BarkerJM HallenbeckDevelopment of an ischemic tolerance model in a PC12 cell lineJ Cereb Blood Flow Metab25154162200510.1038/sj.jcbfm.960000315647748
22.	I SilverM ErecinskaOxygen and ion concentrations in normoxic and hypoxic brain cellsAdv Exp Med Biol454716199810.1007/978-1-4615-4863-8_29889871
23.	SJ RiedlY ShiMolecular mechanisms of caspase regulation during apoptosisNat Rev Mol Cell Biol5897907200410.1038/nrm149615520809
24.	C MoriscoC MarroneV TrimarcoInsulin resistance affects the cytoprotective effect of insulin in cardiomyocytes through an impairment of MAPK phosphatase-1 expressionCardiovasc Res76453464200710.1016/j.cardiores.2007.07.01217698050
25.	G LiotC GabrielM CacquevelNeurotrophin-3-induced PI-3 kinase/Akt signaling rescues cortical neurons from apoptosisExp Neurol1873846200410.1016/j.expneurol.2004.01.00215081586