新型靶向性非病毒基因转移载体K16GRGDSPC介导基因转染的实验研究  

Experimental study on oligopeptide K16GRGDSPC as a targeted vector for gene transfection

作　　者：潘海涛[1] 郑启新[1] 郭晓东[1] 刘勇[1] 李长文[1] 宋玉林[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院骨科,湖北武汉430022

出　　处：《中国医师杂志》2006年第4期433-435,共3页Journal of Chinese Physician

基　　金：国家自然科学基金项目(30170270;30200063及3047083)

摘　　要：目的构建一种新型靶向性非病毒基因转移载体-含RGD肽K16GRGDSPC(记作K16-RGD),评价其介导外源基因转染骨髓基质干细胞的可行性和靶向特异性。方法固相合成法合成K16-RGD,以其为载体将虫荧光素酶质粒导入兔骨髓基质干细胞,检测其转染效率;并观察溶酶体抑制剂氯喹和聚乙烯亚胺(PEI)对其转染效率的影响。通过细胞黏附竞争性抑制和转染竞争性抑制实验检测K16-RGD介导转染的靶向特异性。结果K16-RGD的转染效率低于商业化脂质体,但加入氯喹可使其转染效率与脂质体相近。PEI也能显著提高K16-RGD的转染效率。含RGD肽GRGDSPC、K16GRGDSPC能显著抑制BMSCs的黏附和降低K16-RGD的转染效率。结论K16-RGD是一种易于合成、高效、低毒的新型靶向性非病毒基因转移载体,以其作为载体介导外源基因转染骨髓基质干细胞是可行的,为下一步构建载基因仿生基质材料进行骨缺损修复研究奠定了基础。Objective To investigate the feasibility and efficiency of K16GRGDSPC （K16-RGD） for exogenous gene transfer to bone marrow derived stroma cells （BMSCs）. Methods The peptide K16-RGD was synthesized by solid-phase batch peptide synthesizer. The K16-RGD was used as vector for transfecting the luciferase into BMSCs and the expression of the luciferase gene was monitored. The influence of chloroquine and polyethyleneimine was observed. The targeted specificity was examined by cell attachment test and transfection inhibitation test. Results The transfection efficiency of K16-RGD vector was lower than that of commercial lipofectamine, But the efficency of K16-RGD was greatly enhanced in the presence of chloroquine and polyethyleneimine. The peptides containing RGD inhibited the BMSCs attachment to the 96-well plates and decreased the transfection efficiency of K16-RGD significantly. Conclusion Peptide K16GRGDSPC is a new kind of targeted-nonviral gene delivery vector, which is easy to be synthesized, high efficiency and low cytotoxicity.

关 键 词：基因转移技术 骨髓祖代细胞 甘氨酸 精氨酸 丝氨酸 天冬氨酸 赖氨酸 脯氨酸 半胱氨酸 

分 类 号：R346[医药卫生—基础医学]