军事医学2013，Vol.37Issue(5)：349-353,5.DOI:10.7644/j.issn.1674-9960.2013.05.008
穿膜肽AR-23增强聚乙烯亚胺介导的基因转染效率及其机制的初步研究
Effect of positively charged residues at C-terminal of AR-23 on improvement of polyethylenimine-mediated gene transfection
马倩 1檀英霞 2王颖丽 2李素波 2高红伟 2鲍国强 2季守平2
作者信息
- 1. 中南大学生物科学与技术学院,长沙,410012
- 2. 军事医学科学院野战输血研究所,北京,100850
- 折叠
摘要
Abstract
Objective To investigate the effect of positively charged residues of AR-23 [ a melittin-related peptide ( MRP) from Rana tagoi] on membrane penetrating activity and polyethylenimine ( PEI) -mediated gene transfection. Methods AR-23 and its truncated analog AR-20 (without positively charged residues of AR-23) were synthesized. Then, the second structural information and penetrating ability of peptides were examined by CD spectroscopy and hemolytic activity , respectively. The interaction of peptides with membrane was detected by depolarization study . The effect of peptides on PEI^nediated gene transfection was evaluated using green fluorescence protein ( GFP) and luciferase as reporter genes . The viability of transfected cells was evaluated by MTT assay . Results In the mimetic membrane environment , AR-23 showed typical α-helical structure in which the α-helical content was 37.6% , but AR-20 did not present α-helical conformation. AR-23 exhibited concentration -dependent hemolysis under both neutral and acidic conditions , but AR-20 did not have hemolytic activity. Compared to AR-20, cell membrane was efficiently depolarized by AR -23. Trans fection results showed that luciferase activity and GFP expression in AR -23 /PEI/DNA transfected cells were higher than in PEI /DNA group in both HeLa and RAW264. 7 cells. Compared with PEI transfected group , AR-23 group had higher cell viability in HeLa and RAW264. 7cells. Conclusion The positively charged residues at C-terminal of AR-23 play an important role in maintaining a-helical conformation and membrane penetrating activity . Furthermore, PEI mediated transfection can be effectively improved by AR-23 without causing significant cytotoxicity. Therefore, AR-23 may be considered a potential enhancer of cationic polymers mediated transfection efficiency.关键词
蜂毒肽/穿膜肽/聚乙烯亚胺/膜穿透活性/基因转染Key words
melittin/ cell-penetrating peptides/ polyethylenimine/ membrane penetrating activity/ gene transfection分类
生物科学引用本文复制引用
马倩,檀英霞,王颖丽,李素波,高红伟,鲍国强,季守平..穿膜肽AR-23增强聚乙烯亚胺介导的基因转染效率及其机制的初步研究[J].军事医学,2013,37(5):349-353,5.
