PEG修饰阳离子脂质体对siRNA转染效果的影响

PEG修饰阳离子脂质体对siRNA转染效果的影响

杨丽娟, 杨婷, 姜娟, 徐振中, 王坚成^*, 张强

北京大学药学院药剂学系, 北京 100191

收稿日期:2009-05-07 修回日期:2009-08-10 出版日期:2009-09-15 发布日期:2009-09-15
通讯作者: 王坚成*

Effects of pegylated cationic liposomes on siRNA transfection

Li-Juan Yang, Ting Yang, Juan Jiang, Zhen-Zhong Xu, Jian-Cheng Wang^*, Qiang Zhang

Department of Pharmaceutics, School of Pharmaceutical Sciences, Peking University, Beijing 100191, China

Received:2009-05-07 Revised:2009-08-10 Online:2009-09-15 Published:2009-09-15
Contact: Jian-Cheng Wang*

摘要/Abstract

摘要：

本研究通过体外评价比较了不同阳离子脂材(DC-Chol和DOTAP), 不同比例PEG修饰的阳离子脂质体对siRNA细胞转染效果的影响。体外细胞转染实验结果表明, DOTAP转染能力强于DC-Chol; 2%摩尔比例的PEG可增强细胞内siRNA的摄取。以DOTAP为阳离子脂材的PEG修饰的脂质体与siRNA形成复合物粒径在100 nm左右, 并能减少脂质体与血清中蛋白的相互作用, 从而可能保护siRNA并延长体内循环时间。与阳性转染试剂lipofectamine 2000相比, 该脂质体能有效输送siRNA进入细胞, 并从溶酶体中逃逸。该脂质体可进行进一步的修饰, 有可能应用于siRNA跨膜转运, 并最终用于临床疾病的治疗。

关键词: siRNA, 阳离子脂质体, DC-Chol, DOTAP, PEG

Abstract:

This study aimed to investigate the effects of cationic liposomes containing different cationic lipids (DC-Chol and DOTAP) and different pegylation ratios on siRNA transfection in human U251 glioma cells. The data showed that the transfection efficiency of DOTAP was much higher than that of DC-Chol and PEG at 2 mol% enhanced cellular uptake of siRNA. Cationic liposome-siRNA complexes with particle size around 100 nm were prepared. PEG modification could efficiently stabilize the liposome in the presence of serum, which might protect the siRNA from serum degradation and prolong the circulation time in vivo. Efficient intracellular uptake and lysosome release of siRNA in human U251 glioma cells were observed for pegylated DOTAP-based lipososomes compared with the control transfection reagent lipofectamine 2000. The results demonstrated that this cationic liposome might be a potential vehicle for the in vivo delivery of siRNA.

Key words: siRNA, siRNA, Cationic liposomes, Cationic liposomes, DC-Chol, DC-Chol, DOTAP, DOTAP, PEG, PEG

中图分类号:

R943.4

Supporting:

Abbreviation: DSPE-PEG, methoxypolyethyleneglycol-distearylphospha-tidylethanolamine, PEGMw2000; DC-Chol, 3β[N-(N',N'-dimethyl-aminoethane)carbomyl]cholesterol; DOTAP, {N-[1-(2,3-dioleoyloxy)]-N,N,N-trimethylammoniumpropane}; DOPE, dioleoyl-phosphatidyl-ethanolamine; lipo[C], liposomes composed with DC-Chol/Chol/DOPE; lipo[A], liposomes composed with DOTAP/Chol/ DOPE; PEG-lipo[A], PEG modified liposomes composed with DOTAP/Chol/DOPE/DSPE-PEG.
Foundation items: National Natural Science Foundation of China (Grant No. 30701056), Foundation of MOST (973 Program, Grant No. 2007CB935801), Beijing Natural Science Foundation of China (Grant No. 7083112) and Doctoral Fund of Ministry of Education of China (Grant No. 20070001813).
^*Corresponding author. Tel./fax: 86-10-82802683; e-mail: wang-jc@bjmu.edu.cn

杨丽娟, 杨婷, 姜娟, 徐振中, 王坚成^*, 张强. PEG修饰阳离子脂质体对siRNA转染效果的影响[J]. .

Li-Juan Yang, Ting Yang, Juan Jiang, Zhen-Zhong Xu, Jian-Cheng Wang^*, Qiang Zhang . Effects of pegylated cationic liposomes on siRNA transfection [J]. .

参考文献

[1] Fire, A.; Xu, S.; Montgomery, M.K.; Kostas, S.A.; Driver, S.E.; Mello, C.C. Nature. 1998, 391, 806-811.
[2] Hannon, G.J.; Rossi, J.J. Nature. 2004, 431, 371-378.
[3] Dorsett, Y.; Tuschl, T. Nat. Rev. Drug Discov. 2004, 3, 318-329.
[4] Soutschek, J.; Akinc, A.; Bramlage, B.; Charisse, K.; Constien, R.; Donoghue, M.; Elbashir, S.; Geick, A.; Hadwiger, P.; Harborth, J.; John, M.; Kesavan, V.; Lavine, G.; Pandey, R.K.; Racie, T.; Rajeev, K.G.; Rohl, I.; Toudjarska, I.; Wang, G.; Wuschko, S.; Bumcrot, D.; Koteliansky, V.; Limmer, S.; Manoharan, M.; Vornlocher, H.P. Nature. 2004, 432, 173-178.
[5] Elmen, J.; Thonberg, H.; Ljungberg, K.; Frieden, M.; Westergaard, M.; Xu, Y.; Wahren, B.; Liang, Z.; Orum, H.; Koch, T.; Wahlestedt, C. Nucleic Acids Res. 2005, 33, 439-447.
[6] Chiu, Y.L.; Ali, A.; Chu, C.Y.; Cao, H.; Rana, T.M. Chem. Biol. 2004, 11, 1165-1175.
[7] Braasch, D.A.; Paroo, Z.; Constantinescu, A.; Ren, G.; Oz, O.K.; Mason, R.P.; Corey, D.R. Bioorg. Med. Chem. Lett. 2004, 14, 1139-1143.
[8] Felgner, P.L.; Gadek, T.R.; Holm, M.; Roman, R.; Chan, H.W.; Wenz, M.; Northrop, J.P.; Ringold, G.M.; Danielsen, M. Proc. Natl. Acad. Sci. USA. 1987, 84, 7413-7417.
[9] Malone, R.W.; Felgner, P.L.; Verma, I.M. Proc. Natl. Acad. Sci. USA. 1989, 86, 6077-6081.
[10] Gao, X.; Huang, L. Biochem. Biophys. Res. Commun. 1991, 179, 280-285.
[11] Zelphati, O.; Nguyen, C.; Ferrari, M.; Felgner, J.; Tsai, Y.; Felgner, P.L. Gene Ther. 1998, 5, 1272-1282.
[12] Santel, A.; Aleku, M.; Keil, O.; Endruschat, J.; Esche, V.; Fisch, G.; Dames, S.; Loffler, K.; Fechtner, M.; Arnold, W.; Giese, K.; Klippel, A.; Kaufmann, J. Gene Ther. 2006, 13, 1222-1234.
[13] Li, W.; Szoka, F.C. Jr. Pharm. Res. 2007, 24, 438-449.
[14] Ciani, L.; Ristori, S.; Salvati, A.; Calamai, L.; Martini, G. Biochim. Biophys. Acta. 2004, 1664, 70-79.
[15] Yang, J.P.; Huang, L. Gene Ther. 1997, 4, 950-960.
[16] Li, S.D.; Huang, L. Mol. Pharm. 2006, 3, 579-588.