PDA介导的温和光热联合自噬抑制剂杀伤乳腺癌细胞

PDA-mediated Mild Photothermal Therapy Combined with Autophagy Inhibitors Kill Breast Cancer Cells

目的探究温和光热条件下,抑制自噬是否增强光热治疗的敏感度。方法采用改良的双乳化法和基于聚多巴胺(polydopamine,PDA)的表面修饰法制备CQ@PLGA@PDA NPs;对其进行基本表征后,CCK-8法检测纳米粒子的细胞毒性;近红外激光照射纳米粒子溶液检测升温效果;CCK-8法和活-死细胞染色检测其光热杀伤肿瘤细胞效果;蛋白质免疫印迹实验检测自噬相关蛋白的表达。结果成功制备了CQ@PLGA@PDA NPs,其粒径为253.10±2.39 nm,Zeta电位为-22.57±0.80 mV,粒径均一,分散性好;近红外激光(NIR)辐照纳米粒子溶液10 min后,温度升高至45℃;CQ@PLGA@PDA NPs对细胞无明显毒性,乳腺癌细胞MDA-MB-231和小鼠胚胎成纤维细胞NIH-3T3细胞的存活率均在为95%以上;细胞治疗和蛋白质免疫印迹结果表明,温和光热条件下,抑制自噬可以提高光热治疗的敏感度。结论CQ@PLGA@PDA NPs光热性能好,生物安全性高;通过抑制细胞自噬,可以在温和的光热条件下(<50℃)有效杀伤乳腺肿瘤细胞。

Objective To explore whether inhibiting autophagy can enhance the sensitivity of photothermal treatment under mild photothermal conditions.Methods CQ@PLGA@PDA NPs were prepared by an improved double emulsification method and a PDA-based surface modification method.After basic characterization,CCK-8 method was used to detect the cytotoxicity of nanoparticles;the near-infrared laser irradiation nanoparticle solution was used to detect the heating effect;CCK-8 method and live-dead cell staining were used to detect the killing effect of tumor cells;Western blot was used to detect the expression of autophagy-related proteins.Results The CQ@PLGA@PDA NPs were successfully prepared,with a particle size of 253.10±2.39 nm,a zeta potential of-22.57±0.80 mV,uniform particle size and good dispersion.The temperature of nanoparticle solution increased to 45℃after the near-infrared laser irradiation for 10 min.CQ@PLGA@PDA NPs had no obvious toxicity to cells.The survival rates of breast cancer cell MDA-MB-231 and mouse embryonic fibroblast NIH-3T3 cell were above 95%.The inhibition of autophagy under mild photothermal conditions could improve the sensitivity of photothermal therapy.Conclusion The prepared CQ@PLGA@PDA NPs have good photothermal performance and high biological safety;by inhibiting autophagy,they can effectively kill tumor cells under mild photothermal conditions(<50℃).