Recombinant adenovirus-mediated overexpression of 3β-hydroxysteroid-Δ24 reductase
Xiuli Lu
Dan Jia
Chenguang Zhao
Weiqi Wang
Ting Liu
Shuchao Chen
Xiaoping Quan
Deliang Sun
Bing Gao
摘要：3β-Hydroxysteroid-Δ24 reductase (DHCR24) is a multifunctional enzyme that localizes to the endoplasmic reticulum and has neuroprotective and cholesterol-synthesizing activities. DHCR24 overexpression confers neuroprotection against apoptosis caused by amyloidβdeposition. hTe present study aimed to construct two recombinant adenoviruses driving DHCR24 expression specifically in neurons. Two SYN1 promoter DNA fragments were obtained from human (h) and rat (r). Recombinant Ad-r(h)SYN1-DHCR24 was transfected into AD-293, N2A (mouse neuroblastoma), and MIN6 (mouse pancreatic carcinoma) cells. Western blot analysis showed DHCR24 was specially expressed in 293 and N2A cells, but no speciifc band was found in MIN6 cells. hTis demonstrates that the recombinant adenoviruses successfully express DHCR24, and no expression is observed in non-neuronal cells. TUNEL assay results showed apoptosis was inhibited in adenovirus-transfected neurons. Detecting reactive oxygen species by immunolfu-orescence, we found that adenovirus transfection inhibits apoptosis through scavenging excess reactive oxygen species. Our ifndings show that the recombinant DHCR24 adenoviruses induce neuron-specific DHCR24 expression, and thereby lay the foundation for further studies on DHCR24 gene therapy for Alzheimer’s disease.
机标关键词：recombinant adenovirusesreactive oxygen speciesendoplasmic reticulumpancreatic carcinomagene therapy
资助基金：Excellent Talent Support Program of Liaoning Province, (No. LJQ2011004)
论文发表日期：2014-01-01
在线出版日期：2025-08-15（本平台首次上网日期，不代表文献的发表时间）
页数：9( 504-512 )
英文信息