偶联双基因靶点的多功能金纳米颗粒对三阴性乳腺癌细胞的毒性分析

摘要

目的：：设计并构建偶联双基因靶点的多功能金纳米棒颗粒(TKI-258-pGNRs@mSiO2-RGD),并探讨该多功能金纳米棒颗粒对人三阴性乳腺癌MDA-MB-231细胞的细胞毒性,初步明确该新型多功能金纳米棒颗粒在体外实验中所需的剂量范围。方法：运用种子介导生长法制备金纳米棒,以改良的奥伯法对其表面进行介孔二氧化硅包裹,通过表面修饰偶联整合素αvβ3和表皮生长因子受体( VEGFR )的特异性抑制剂RGD和TKI-258,采用紫外分光光度计和透射电镜对其特征进行检测。以不同浓度(0、12.5、25、50、100、200μg/ml)多功能金纳米棒颗粒孵育人三阴性乳腺癌MDA-MB-231细胞株,采用CCK-8法检测细胞增殖率,并进行细胞毒性评级。结果：多功能金纳米棒颗粒物理状态稳定,吸收峰位于515、860 nm处。偶联的双基因靶点抑制剂TKI-258、RGD分别在360、204 nm 处有特征性吸收峰,药物包封率分别为58.18%±7.13%,57.95%±5.22%,载药率分别为4.26%±0.42%,4.35%±0.79%。不同浓度多功能金纳米棒处理后,细胞增殖率在50%-100%之间,细胞毒性评级为I-II级,0-100μg/ml浓度范围内无细胞毒性。结论：偶联双基因靶点的多功能金纳米棒颗粒( TKI-258-pGNRs@mSiO2-RGD)具有热疗/放疗增敏的特殊光学性质,偶联双基因靶点抑制剂TKI-258、RGD的载药量稳定,生物相容性良好,可作为三阴性乳腺癌细胞特异性靶向的热疗/放疗载药增敏材料。%Objective:Background and objective:The combination of nanotechnology and phototherapy/radiothera-py is a new focus in tumor therapy. Our study design and synthesized multifunctional gold nanoparticles conjugated with dual-target inhibitors( TKI-258-pGNRs@mSiO2-RGD) andanalysis cytotoxicity on human triple negative breast cancer MDA-MB-231 cell lineto preliminary identify the effective dose of multifunctional gold nanoparticles in vitro. Methods:Gold nanorods were synthesized byseed-mediated growth method and coated with mesoporous silica by modified Stober method. The integrinαvβ3 and vascular endothelial growth factor receptor ( VEGFR) inhibitors RGD and TKI-258 were conjugated on goldnanorods. The physical properties of gold nanorods were characterized by trans-mission electron microscope and UV-vis spectrometer respectively. The multifunctional nanorods were prepared in so-lution at different concentrations(0,12. 5,25,50,100,200μg/ml) and were incubated by MDA-MB-231 cell line, cell proliferation and cytotoxicity grading were detected by the cell counting kit-8 ( CCK-8 ) method. Results:The multifunctional gold nanorods had steady physical property and its absorption peaks at 515nm and 860nm,respective-ly. The dual-target inhibitors TKI-258 and RGD had special absorption peaks at 360nm and 204nm,respectively. En-capsulationefficiencywere58.18%±7.13% and57.95%±5.22%,respectively.Drugloadingratiowere4.26%±0. 42% and 4. 35%±0. 79%,respectively. After co-cultured with the multifunctional gold nanorods on different concentration,relative growth rate of MDA-MB-231 cell line was between 50% and 100%,cytotoxicity grading was level I -II. There was no cytotoxicity on the concentrations of 0-100μg/ml. Conclusion:The multifunctional gold nanorads conjugated with dual-target inhibitors ( TKI-258-pGNRs@mSiO2-RGD) had special optical properties on phototherapy/radiotherapyand stable drug loading capacity. It's aphototherapy/radiotherapysensitizing agentswith well biocompatibility targeted on human triple negative breast cancer MDA-MB-231 cell line.