首页|期刊导航|中国神经再生研究（英文版）|Adenovirus-mediated short hairpin RNA interference against p75 neurotrophin receptor in pheochromocytoma cells
中国神经再生研究（英文版）2011，Vol.6Issue(7)：517-522,6.DOI:10.3969/j.issn.1673-5374.2011.07.006
Adenovirus-mediated short hairpin RNA interference against p75 neurotrophin receptor in pheochromocytoma cells
Adenovirus-mediated short hairpin RNA interference against p75 neurotrophin receptor in pheochromocytoma cells
摘要
Abstract
Previous studies have confirmed that motor neuron apoptosis in the anterior horn of the lumbosacral spinal cord is positively correlated with p75 neurotrophin receptor (p75NTR) expression in rat models of cauda equina syndrome. This study used adenovirus to carry a short hairpin RNA (shRNA) for p75NTR gene silencing, to reduce p75NTR expression in the damaged phase and to decrease motor neuron apoptosis. Three p75 siRNA template oligonucleotide segments (shRNA) were designed, and cloned into the 1.0 CMV shuttle vector. HEK293 cells were cotransfected with shuttle vector (carrying shRNA) and an adenovirus vector framework expressing enhanced green fluorescent protein. Thus, this study successfully obtained adenovirus carrying p75shRNA. The obtained viruses were named Ad.shRNA1, Ad.shRNA2, and Ad.shRNA3. The recombinant adenoviruses were separately used to infect cultured pheochromocytoma cells (PC12). Forty-eight hours later, p75NTR mRNA and total protein were analyzed from the PC12 cells. Compared with the negative controls, RNA interference rates were separately 98.49±0.68%, 95.08±1.79% and 96.60±1.14% at the mRNA level, and 72.89±2.17%, 58.83±1.15% and 59.88±0.44% at the protein level in the Ad.shRNA1, Ad.shRNA2, and Ad.shRNA3 groups, respectively. Thus, recombinant adenovirus shRNA-mediated gene silencing successfully suppressed p75NTR expression.关键词
p75 neurotrophin receptor/RNA interference/adenovirus/rat pheochromocytoma cells/human embryonic kidney 293 cells/apoptosis/cauda equina syndromeKey words
p75 neurotrophin receptor/RNA interference/adenovirus/rat pheochromocytoma cells/human embryonic kidney 293 cells/apoptosis/cauda equina syndrome引用本文复制引用
Dongxu Feng,Haopeng Li,Siyue Xu,Yu Liu,Xiaofei Hou..Adenovirus-mediated short hairpin RNA interference against p75 neurotrophin receptor in pheochromocytoma cells[J].中国神经再生研究（英文版）,2011,6(7):517-522,6.基金项目
the National Natural Science Foundation of China,No.30672136 （）
