FeSe_(2)纳米粒子对膀胱癌T24细胞的抑制作用及其机制

INHIBITORY EFFECT OF FESE_(2) NANOPARTICLES ON BLADDER CANCER T24 CELLS AND ITS MECHANISM

目的制备一种可降解谷胱甘肽(GSH)、升高细胞内活性氧(ROS)的FeSe_(2)纳米粒子,并探究其对膀胱癌T24细胞的抑制作用。方法以乙醇胺为溶剂,用氯化亚铁和硒粉一步水热法合成FeSe_(2)纳米粒子,对FeSe_(2)进行透射电镜、X射线衍射表征;采用噻唑蓝(MTT)法检测FeSe_(2)纳米粒子对T24细胞和人正常膀胱上皮SV-HUC-1细胞活性的影响;采用5,5′-二硫代双(2-硝基苯甲酸)(DTNB)法检测FeSe_(2)在试管中与GSH共孵育后GSH含量变化;采用4′,6-二脒基-2-苯基吲哚(DAPI)与2′,7′-二氯二氢荧光素二乙酸酯(DCFH-DA)双染检测T24细胞内ROS水平;采用MTT法检测铁死亡抑制剂Ferrostatin-1干预后,FeSe_(2)对T24细胞活性的影响。结果成功合成了直径10~20 nm的FeSe_(2)纳米粒子,该粒子对T24细胞活性的抑制作用强于对SV-HUC-1细胞活性的抑制作用(F=868.60,P<0.05);FeSe_(2)在试管中与GSH共孵育后可降低GSH的含量,并可升高T24细胞内ROS的水平;Ferrostatin-1可逆转FeSe_(2)对膀胱癌T24细胞的抑制作用。结论本研究采用一步水热法成功合成了粒径小、结晶度高的FeSe_(2)粒子,该粒子可通过降解胞外GSH,升高胞内ROS水平,实现对膀胱癌T24细胞活性的抑制作用。

Objective To prepare FeSe_(2) nanoparticles that can degrade glutathione(GSH)and increase intracellular reactive oxygen species(ROS),and to investigate its inhibitory effect on bladder cancer T24 cells.Methods FeSe_(2) nanoparticles were synthesized by the one-step hydrothermal method with ferrous chloride and selenium powder using ethanolamine as the solvent and then FeSe_(2) was characterized by transmission electron microscopy and X-ray diffraction.MTT assay was used to observe the effect of FeSe_(2)on the viability of T24 cells and human normal bladder epithelial SV-HUC-1 cells.The 5,5′-Dithiobis(2-nitrobenzoic acid)method was used to measure the change in the content of GSH after co-incubation of FeSe_(2)and GSH in vitro.Double staining with 4′,6-diamidino-2-phenylindole and 2′,7′-dichlorodihydrofluorescein diacetate was used to measure the level of ROS in T24 cells.MTT assay was used to observe the effect of FeSe_(2)on the viability of T24 cells after intervention with the ferroptosis inhibitor Ferrostatin-1.Results FeSe_(2)nanoparticles with a diameter of 10-20 nm were successfully synthesized,and the inhibitory effect of FeSe_(2)nanoparticles on T24 cell viability was stronger than that on SV-HUC-1 cell viability(F=868.60,P<0.05).After co-incubation with GSH in vitro,FeSe_(2)reduced the content of GSH and increased the level of ROS in T24 cells,and Ferrostatin-1 reversed the inhibitory effect of FeSe_(2)on T24 cells.Conclusion This study uses the one-step hydrothermal method to successfully synthesize FeSe_(2)particles with small particle size and high crystallinity,which exerts an inhibitory effect on the viability of T24 cells by degrading extracellular GSH and increasing the level of intracellular ROS.