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  

Cell-penetrating chimeric apoptotic peptide AVPI-LMWP/DNA co-delivery system for cancer therapy

作　　者：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] 

机构地区：[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

出　　处：《药学学报》2014年第12期1718-1723,共6页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81373357;81422048;81172996)

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/p TRAIL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(p TRAIL)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/p TRAIL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/p TRAIL复合物,随着AVPI-LMWP/p TRAIL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p TRAIL。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/p TRAIL(W∶W=20∶1)复合物抑制He La细胞增殖的效果显著强于p TRAIL、AVPI-LMWP及LMWP/p TARIL复合物。研究显示,AVPI-LMWP/p TRAIL基因共给药系统能将质粒高效递送到He La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。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 was improved along with the increased ratio of demonstrated that the AVPI-LMWP/pTRAIL （W： at a mass ratio above 15 ： 1. Cellular uptake efficiency WAVPI-LMWP/WpTRAIL. The in vitro cytotoxicity experiments W=20 ： 1） complexes was significantly more effective than the pTRAIL, AVPI-LMWP alone or LMWP/pTRA1L 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.

关 键 词：穿膜肽 凋亡肽 基因给药 细胞凋亡 TRAIL AVPI 肿瘤治疗 

分 类 号：R943[医药卫生—药剂学]