药学学报, 2014, 49(12): 1718-1723
引用本文:
谭娇, 王雅萍, 王慧欣, 梁剑铭, 张梦, 孙逊, 黄永焯. 具有穿膜功能的嵌合型AVPI-低分子量鱼精蛋白/DNA共给药系统的抗肿瘤研究[J]. 药学学报, 2014, 49(12): 1718-1723.
TAN Jiao, WANG Ya-ping, WANG Hui-xin, LIANG Jian-ming, ZHANG Meng, SUN Xun, HUANG Yong-zhuo. Cell-penetrating chimeric apoptotic peptide AVPI-LMWP/DNA co-delivery system for cancer therapy[J]. Acta Pharmaceutica Sinica, 2014, 49(12): 1718-1723.

具有穿膜功能的嵌合型AVPI-低分子量鱼精蛋白/DNA共给药系统的抗肿瘤研究
谭娇1,2, 王雅萍2, 王慧欣2, 梁剑铭2, 张梦2, 孙逊1, 黄永焯2
1. 四川大学华西药学院, 四川 成都 610041;
2. 中国科学院上海药物研究所, 上海 201203
摘要:
本文旨在合成具有穿膜活性的促凋亡AVPI-低分子量鱼精蛋白嵌合多肽 (AVPI-LMWP), 制备安全有效的AVPI-LMWP/pTRAIL基因共给药系统, 通过凋亡肽AVPI与肿瘤坏死因子相关凋亡诱导配体基因 (pTRAIL) 联合用药来发挥抗肿瘤作用.采用标准固相合成法合成嵌合型多肽AVPI-LMWP.基于静电吸附作用, AVPI-LMWP可与DNA药物形成AVPI-LMWP/pTRAIL 复合物, 并对其理化性质进行了表征, 同时研究其细胞摄取效率和抗肿瘤增殖能力.结果表明, 采用共孵育法成功制备了AVPI-LMWP/pTRAIL复合物, 随着AVPI-LMWP/pTRAIL质量比的增加, 复合物的平均粒径逐渐减小, zeta-电位变化较小.凝胶电泳实验结果表明, 当两者质量比大于15:1时, AVPI-LMWP能完全包裹和压缩质粒pTRAIL.体外细胞摄取实验显示, 随着二者质量比的增加, 细胞摄取效率有所提高.体外细胞毒性实验表明, AVPI-LMWP/pTRAIL (W:W = 20:1) 复合物抑制HeLa细胞增殖的效果显著强于pTRAIL、AVPI-LMWP及LMWP/pTARIL复合物.研究显示, AVPI-LMWP/pTRAIL基因共给药系统能将质粒高效递送到HeLa细胞内, 并能有效地诱导肿瘤细胞凋亡, 是具有联合治疗应用价值的多肽/基因共给药系统.
关键词:    穿膜肽      凋亡肽      基因给药      细胞凋亡      TRAIL      AVPI      肿瘤治疗     
Cell-penetrating chimeric apoptotic peptide AVPI-LMWP/DNA co-delivery system for cancer therapy
TAN Jiao1,2, WANG Ya-ping2, WANG Hui-xin2, LIANG Jian-ming2, ZHANG Meng2, SUN Xun1, HUANG Yong-zhuo2
1. West China School of Pharmacy, Sichuan University, Chengdu 610041, China;
2. Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China
Abstract:
To develop a cell-penetrating chimeric apoptotic peptide AVPI-LMWP/DNA co-delivery system for cancer therapy, we prepared the AVPI-LMWP/pTRAIL self-assembled complexes containing a therapeutic combination of peptide drug AVPI and DNA drug TRAIL. The chimeric apoptotic peptide AVPI-LMWP was synthesized using the standard solid-phase synthesis. The cationic AVPI-LMWP could condense pTRAIL by electrostatic interaction. The physical-chemical properties of the AVPI-LMWP/pTRAIL complexes were characterized. The cellular uptake efficiency and the inhibitory activity of the AVPI-LMWP/pTRAIL complexes on tumor cell were also performed. The results showed that the AVPI-LMWP/pTRAIL complexes were successfully prepared by co-incubation. With the increase of mass ratio (AVPI-LMWP/DNA), the particle size was decreased and the zeta potential had few change. Agarose gel electrophoresis showed that AVPI-LMWP could fully bind and condense pTRAIL at a mass ratio above 15:1. Cellular uptake efficiency was improved along with the increased ratio of WAVPI-LMWP/WpTRAIL. The in vitro cytotoxicity experiments demonstrated that the AVPI-LMWP/pTRAIL (W:W = 20:1) complexes was significantly more effective than the pTRAIL, AVPI-LMWP alone or LMWP/pTRAIL complexes on inhibition of HeLa cell growth. Our studies indicated that the AVPI-LMWP/pTRAIL co-delivery system could deliver plasmid into HeLa cell and induce tumor cell apoptosis efficiently, which showed its potential in cancer therapy using combination of apoptoic peptide and gene drugs.
Key words:    cell penetrating peptide    apoptotic peptide    gene delivery    apoptosis    TRAIL    AVPI    cancer therapy   
收稿日期: 2014-04-15
基金项目: 国家自然科学基金资助项目(81373357,81422048,81172996).
通讯作者: 孙逊, 黄永焯
Email: xunsun22@gmail.com;yzhuang@mail.shcnc.ac.cn
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