纳米化色胺酮类化合物CY-1-4诱导B16-F10细胞发生铁死亡

Tryptanthrin derivative CY-1-4 nanoparticle induces ferroptosis in B16-F10 cells

CY-1-4是一种色胺酮类化合物,其抗肿瘤作用已被证实,但其水溶性差,且抗肿瘤作用机制尚未阐明。针对这些问题,本研究首先以聚己内酯[poly(caprolactone),PCL]和聚乙二醇-聚ε-己内酯[poly(ethylene glycol)-copoly(ε-caprolactone),PEG-PCL]为载体材料,通过纳米沉淀法制备了包载CY-1-4的纳米粒(CY-1-4 NPs)以改进其溶解性,测定CY-1-4 NPs对B16-F10细胞活性氧(reactive oxygen species,ROS)水平的影响、脂质活性氧抑制剂ferrostatin-1和铁螯合剂去铁胺(deferoxamine,DFO)对CY-1-4 NPs诱导B16-F10细胞死亡的修复作用及原卟啉(protoporphyrin IX,PPIX)对CY-1-4 NPs诱导B16-F10细胞死亡的促进作用,研究了CY-1-4 NPs诱导B16-F10的细胞毒是否存在铁死亡途径。结果表明,纳米化策略可明显改善CY-1-4的水溶性,所得纳米粒粒径约为116 nm,包封率约为83%,载药量约为4.80%。铁死亡机制验证结果表明,CY-1-4 NPs可以明显提高B16-F10细胞内ROS水平,ferrostatin-1和DFO可以一定程度抑制CY-1-4 NPs对B16-F10细胞的细胞毒作用,而PPIX可以促进CY-1-4 NPs对B16-F10细胞的细胞毒。上述结果均证明铁死亡是纳米化色胺酮类化合物CY-1-4诱导细胞死亡的机制之一。

CY-1-4 is a tryptanthrin derivative exhibiting antitumor activity.The solubility of CY-1-4 was poor and the corresponding mechanism needs further study.To solve this problem,we prepared nanoparticles encapsulated with CY-1-4(CY-1-4 NPs) by nanoprecipitation method using poly(caprolactone)(PCL) and poly(ethylene glycol)-co-poly(ε-caprolactone)(PEG-PCL) as carriers to improve solubility.We then explored whether CY-1-4 NPs induced B16-F10 cytotoxicity via ferroptosis by determining the effect of CY-1-4 NPs on reactive oxygen(ROS)levels,repairing efficacy of lipid reactive oxygen inhibitor ferrostatin-1 and iron chelator deferoxamine(DFO),and potentiation of protoporphyrin(PPIX) induced B16-F10 cell death.The results showed that nanoparticlated strategy significantly improved solubility of CY-1-4.With the particle size about 116 nm,encapsulating efficacy was about 83% and the drug loading capacity was about 4.80%.Ferroptosis mechanistic studies indicated that CY-1-4 NPs could improve the ROS level in B16-F10 cells,whereas ferrostatin-1 and DFO could partly inhibited the cytotoxicity and PPIX could potentiated the cytotoxicity of CY-1-4 NPs in B16-F10 cells.These results showed that ferroptosis was one of the cell death mechanisms induced by tryptanthrin derivative CY-1-4 nanoparticle.