白细胞介素15基因脂质体的构建及其抗肿瘤效果评价

Study on the preparation and the anti-tumor effect evaluation of pIL-15 liposomes

目的制备DSPE-PEG2000-FA修饰的阳离子脂质体(FPCL),并与白细胞介素15质粒DNA(interleukin-15 plasmid DNA,IL-15 pcDNA,pIL-15)形成FPCL/pIL-15基因递送系统,并进行抗肿瘤效果评价。方法通过薄膜分散法制备FPCL,并且利用正负电荷吸引的原理,制备FPCL/pIL-15复合物。运用动态光散射法测定复合物粒径,利用琼脂糖凝胶电泳考察复合物稳定性,采用荧光分光光度法测定其包封率,以Balb/c小鼠为模型,考察制剂的抗肿瘤效果。结果成功制备了FPCL/pIL-15阳离子脂质体基因复合物,FPCL/pDNA复合物在质量比w_((DOTAP))∶w_((pDNA))=5∶1时粒径达到200 nm以下;抵御阴离子置换实验表明FPCL包载pDNA的稳定性良好,药效学实验表明FPCL/pIL-15复合物抗肿瘤效果显著。结论FPCL作为基因递送载体对基因药物具有很好的保护性,FPCL/pIL-15复合物能够有效诱导NK-细胞增殖与激活,抑瘤效果明显,在肿瘤免疫治疗领域有很大的研究价值。

Objective To prepare cationic liposomes modified by DSPE-PEG2000-FA(FPCL)for the formation of FPCL/pIL-15 gene delivery system with interleukin-15 plasmid DNA(pIL-15)and to evaluate and discuss the anti-tumor effect.Methods FPCL prepared by the membrane dispersion method,was utilized to prepare positive cationic liposome/pIL-15 complex based on the principle of positive and negative charge attraction.The particle size,stability and encapsulation efficiency were determined by dynamic light scattering method,agarose gel electrophoresis and fluorescence spectrophotometry,respectively.The antitumor effect of the preparation was investigated using Balb/c mice model.Results Positive cationic liposome/pIL-15 complexes were prepared successfully.When the mass ratio of FPCL/pIL-15 was 5∶1(w_((DOTAP))∶w_((pDNA))=5∶1),the diameter of particle was below 200 nm.And with the increase of mass ratio,the particle size changed slowly and tended to be stable eventually.The resistance to anion replacement experiment showed that the stability of FPCL/pDNA was admirable.FPCL/pDNA had the significant antitumor effect.Conclusion As a gene delivery vector,FPCL has the potent ability of protecting gene drugs compared with PCL.FPCL/pil-15 complex can effectively induce the proliferation and activation of NK cells,and has obvious anti-tumor effect,which shows promising future in the field of tumor immunotherapy.