VEGF修饰的载TNP-470纳米粒对荷瘤裸鼠的靶向性研究

Study on the tumor targeting of VEGF modified nanoparticles containing TNP-470 in tumor-bearing nude mouse models

目的:制备一种具有靶向新生血管特点的载TNP-470纳米粒,观察其对人脐静脉内皮细胞(HUVECs)的靶向性及增殖的影响,观察其对肿瘤组织的靶向性以及治疗效果。方法:采用单乳法制备载TNP-470纳米粒,并将其共价偶联血管内皮生长因子(VEGF),命名为TNP-470-NPs-VEGF。透射电镜下观察TNP-470-NPs-VEGF的形态,纳米粒度分析仪检测其大小。CCK-8试剂盒检测对HUVECs的细胞毒作用,荧光显微镜观察HUVECs对纳米粒的摄取。观察静脉注射后对肿瘤的靶向性,以及对荷瘤裸鼠的治疗效果。结果:TNP-470-NPs-VEGF形态近似球形,直径为115～130nm。细胞摄取结果显示,VEGF修饰的纳米粒能明显被细胞所摄取,且呈浓度依赖性;CCK-8结果显示,VEGF修饰的载TNP-470靶向纳米粒能够明显抑制HUVECs细胞的生长,且呈浓度依赖性。SKOV3荷瘤裸鼠体内实验发现,TNP-470-NPs-VEGF的肿瘤靶向性好,能显著抑制肿瘤的生长。结论:成功制备得到的TNP-470-NPs-VEGF,能显著增强HUVECs细胞的摄取,抑制细胞增殖,对裸鼠体内的人卵巢癌SKOV3肿瘤具有明显的靶向性,能显著抑制肿瘤的生长。

Objective：To prepare an angiogenesis-targeting TNP-470-loaded nanoparticles（TNP-470-NPs） and evaluate its effects on human umbilieal vein endothelial cells（HUVECs）.In addition,to analyze its targeting capability and therapeutic effects with tumor tissues.Methods：TNP-470-NPs was prepared by emulsion/solvent evaporation method and conjugated with vascular endothelial growth factor（VEGF）,then was named for TNP-470-NPs-VEGF.The morphology of the NPs was observed by transmission electron microscopy（TEM） and the particle size was determined by nanoparticle size analyzer.Its effects on proliferation were measured by cell counting kit-8（CCK-8）.The cell uptake was observed by inverted microscope.The targeting capability and therapeutic effects were evaluated in tumor-bearing nude mouse models.Results：TNP-470-NPs-VEGF was spherical with an average diameter 131.25 nm.The results of cell uptake and proliferation indicated that TNP-470-NPs-VEGF can significantly inhibit the proliferation and increase the cell uptake of HUVECs in dose-dependent manner.TNP-470-NPs-VEGF had obvious tumor-targeting effects and can significantly inhibit the growth of tumor on human ovarian cancer SKOV3 nude mice.Conclusion：Success in the preparation of TNP-470-NPs-VEGF,which significantly inhibited HUVECs migration and had considerable tumor-targeting and antitumor effects on human ovarian cancer SKOV3 nude mice.