二茂铁纳米平台逆转索拉非尼化疗耐药性的研究

Ferrocene nanoplatforms reverse sorafenib chemoresistance

目的研究制备的负载索拉非尼(Sorafenib,SFB)的二茂铁金属有机骨架(MOF-Fc)纳米平台(SFB@MOF-Fc),逆转肿瘤耐药性的机制与效果。方法通过流式细胞术以及细胞存活率实验,考察SFB耐药肿瘤细胞模型的构建及耐药机制。通过细胞存活率以及3D肿瘤球的细胞活/死染色实验,考察纳米平台体外抗肿瘤能力。通过检测细胞内活性氧(Reactive Oxygen Species,ROS)水平以及生物电镜研究,考察纳米平台体外抗肿瘤机制。结果通过对细胞存活率分析,显示出SFB耐药细胞(Huh7-SR)的成功建立。通过对流式细胞术检测ROS水平分析,显示Huh7/SR细胞在被SFB药物处理后,ROS水平升高不明显,表明ROS水平可能是肿瘤耐药的关键因素。通过细胞存活率以及3D肿瘤球细胞活/死染色分析,显示SFB@MOF-Fc组处理后,细胞存活率降低以及死细胞荧光染色区域增加,表明SFB@MOF-Fc成功逆转了体外模型的耐药性。通过ROS荧光探针以及生物电镜分析,表明SFB@MOF-Fc纳米粒可以通过使Huh7-SR细胞产生铁死亡,放大氧化应激水平的机制来逆转肿瘤耐药。结论SFB@MOF-Fc被肿瘤细胞内吞后,药物在细胞的摄取增加,通过铁死亡,放大氧化应激水平,达到了有效逆转肿瘤细胞耐药的效果。

Objective To investigate the mechanism and effect of nanocomposites(SFB@MOF-Fc)loaded with sorafenib(SFB)to reverse tumor drug resistance.Methods The construction of SFB-resistant tumor cell model and the study of resistance mechanism were investigated by flow cytometry and cell viability.The in vitro anti-tumor ability of drug-carrying nanoplatforms was investigated by cell viability and 3D tumor sphere cell live/dead staining analysis.The in vitro anti-tumor mechanism of drug-carrying nanoplatforms was investigated by the detection of the level of intracellular reactive oxygen species(ROS)and by bioelectron microscopy study.Results SFB-resistant cells(Huh7-SR)showed successful establishment by the cell viability analysis.The analysis of ROS levels in Huh7 cells as well as Huh7-SR cells detected by the flow cytometry showed that ROS levels in Huh7-SR were insignificantly elevation after SFB drug treatment,suggesting that ROS levels may influence resistance mechanisms.The analysis of cell viability as well as 3D tumorsphere cell live/dead staining showed that the cell viability as well as cell death fluorescence were greatly increased after SFB@MOF-Fc treatment,indicating the SFB@MOF-Fc successfully reversed drug resistance in the in vitro model.ROS levels as well as bioelectron microscopy analyses showed that SFB@MOF-Fc composite nanomaterials could reverse the tumor resistance through a mechanism that enhances the production of iron death in Huh7-SR cells and amplifies the level of oxidative stress.Conclusion After SFB@MOF-Fc is endocytosed by tumor cells,the drug uptake in cells increased.The oxidative stress levels were amplified through intracellular ferroptosis,and successfully reversed the drug resistance of tumor cells.