RGD修饰的低分子量鱼精蛋白/CpG ODN复合物的制备及抗肿瘤活性  

Preparation and Antitumor Activity of RGD-modified Low Molecular Weight Protamine/CpG complexes

作　　者：刘佳佳 苏秋东[1] 伊瑶[1] 毕胜利[1] LIU Jiajia;SU Qiudong;YI Yao(National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China)

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京102206

出　　处：《医学研究杂志》2023年第1期130-135,共6页Journal of Medical Research

摘　　要：目的利用RGD修饰的低分子量鱼精蛋白(low molecular weight protamine,LMWP)包裹CpG寡聚脱氧核苷酸(CpG oligodeoxynucleotide,CpG ODN)制备复合物,评价其对CpG ODN抗肿瘤活性的影响。方法基于静电引力将RGD-LMWP与CpG ODN以不同质量比共孵育制备复合物,并利用凝胶阻滞实验分析RGD-LMWP对CpG的缩合能力;使用zeta电位/粒度仪对RGD-LMWP/CpG复合物的粒径和zeta电位进行表征;采用体外细胞实验评价RGD-LMWP/CpG复合物对肿瘤细胞的摄取效率和细胞毒性。结果采用共孵育法成功制备了RGD-LMWP/CpG复合物。当RGD-LMWP与CpG的质量比为3∶1时,RGD-LMWP能完全压缩CpG。此时,复合物的平均粒径和zeta电位分别为119.7±1.2nm和47.9±0.5mV。体外细胞实验表明,与游离CpG比较,RGD-LMWP/CpG(3∶1)复合物能显著提高人子宫颈癌细胞株(HeLa)细胞对CpG的摄取效率,抑制HeLa细胞的增殖。结论RGD-LMWP与CpG结合形成的复合物可以有效地将CpG递送至肿瘤细胞内并诱导肿瘤细胞凋亡,是一种具有研究前景的抗肿瘤给药方案。Objective CpG oligodeoxynucleotide(CpG ODN)was coated with low molecular weight protamine(LMWP)modified by RGD to prepare complex,to evaluate its effect on the antitumor activity of CpG ODN.Methods RGD-LMWP/CpG complexes were prepared by coincubating RGD-LMWP peptide and CpG at different mass ratios based on electrostatic attraction,and the condensation ability of RGD-LMWP to CpG was analyzed by gel block experiment.The particle size and zeta potential of the RGD-LMWP/CpG complexes were characterized by zeta potential/particle size analyzer.In addition,the uptake efficiency and cytotoxicity of RGD-LMWP/CpG complexes to tumor cells were also examined by in vitro cell assay.Results The RGD-LMWP/CpG complexes were successfully prepared by coincubating RGD-LMWP and CpG.When the mass ratio of RGD-LMWP to CpG was increased to 3∶1,RGD-LMWP could completely condense CpG.The average particle size and zeta potential of RGD-LMWP/CpG complexes were 119.7±1.2nm and 47.9±0.5mV,respectively,which were suitable for cellular uptake.In vitro cell experiments showed that compared with the free CpG,RGD-LMWP/CpG(3∶1)complexes could significantly improve the uptake efficiency of CpG by HeLa cells,thereby inhibited the proliferation of HeLa cells.Conclusion RGD-LMWP/CpG complexes can efficiently deliver CpG into tumor cells and induce tumor cell apoptosis,which is a promising candidate for antitumor therapy.

关 键 词：低分子量鱼精蛋白 RGD CPG寡聚脱氧核苷酸 抗肿瘤活性 

分 类 号：R979.1[医药卫生—药品]