中国药理学通报2016，Vol.32Issue(8)：1153-1157,1158,6.DOI:10.3969/j.issn.1001-1978.2016.08.024
慢病毒介导的 NGF基因沉默对 PC12细胞分化的影响
Lentivirus-mediated NGF gene silencing inhibited differentiation of PC12 Cells
摘要
Abstract
Aim To investigate the effects of lentivirus mediated nerve growth factor ( NGF) gene silencing on pheochromocytoma cells ( PC12 ) and the possible mechanisms .Methods The NGF shRNA expression vector was constructed .PC12 cells were randomly divi-ded into five groups (n=3 each) as follows: negative control group ( NC ) , control lentivirus group ( LV CON) , lentivirus NGF shRNA1 group ( LV shNGF1 ) , lentivirus NGF shRNA2 group(LV shNGF2), lentivir-us NGF shRNA3 group(LV shNGF3).The cells in NC group were cultured in DMEM/HG and polybrene me-dium, while others were cultured in DMEM/HG, poly-brene and corresponding lentivirus medium .After the treatment, the infection efficiency was determined by fluorescent microscope .Relative expression of NGF , extracellular signal-regulated kinase ( ERK1/2 ) and p-ERK1/2 were assessed by Western blot .The expres-sion of NGF mRNA was analyzed by quantitative re-verse transcription polymerase chain reaction ( qRT-PCR) .The differentiation degree was valued according to the length of neuritis and max diameter of cells .The cell viability was detected by CCK-8.Results The in-fection efficiency in PC12 cells reached over 90%. Compared with NC group , the relative expression of NGF mRNA and NGF protein was significantly down-regulated ( P<0.05 ) .There was no difference in the expression of ERK1/2 protein and cell viability .The expression of p-ERK1/2 protein was markedly down-regulated in LV shNGF3 group ( P<0.01 ) .The cells morphology was changed , and the length of neuritis and max diameter of cells were strained in LV shNGF 3 group than those in NC group ( P<0.01 ) .Conclusion Lentivirus-mediated NGF gene silencing inhibits the differentiation of PC12 cells through suppressing the activation of ERK1/2.关键词
慢病毒/NGF/基因沉默/PC12细胞/分化/ERK1/2Key words
lentivirus/NGF/gene silencing/pheo-chromocytoma cell/differentiation/extracellular signal-regulated kinase分类
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窦梦云,何淑芳,黄成,潘永露,张野..慢病毒介导的 NGF基因沉默对 PC12细胞分化的影响[J].中国药理学通报,2016,32(8):1153-1157,1158,6.基金项目
国家自然科学基金资助项目 （）
