卡非佐米联合碘-125粒子照射促进人食管癌细胞KYSE-150凋亡的机制研究  

The pro-apoptotic effect of carfilzomib combined with iodine-125 seed irradiation on KYSE-150 esophageal cancer cells

作　　者：王超[1] 王浩 孙柏 袁野 羌伟光 石红兵[1] Chao Wang;Hao Wang;Bai Sun;Ye Yuan;Weiguang Qiang;Hongbing Shi(Department of Oncology,the First People's Hospital of Changzhou,Jiangsu Changzhou 213003,China)

机构地区：[1]常州市第一人民医院肿瘤科,江苏常州213003

出　　处：《中华介入放射学电子杂志》2024年第2期106-113,共8页Chinese Journal of Interventional Radiology：electronic edition

基　　金：江苏省自然科学基金项目(BK20210085)。

摘　　要：目的探索卡非佐米(carfilzomib,CFZ)联合碘-125(iodine-125,^(125)I)粒子照射对人食管癌细胞KYSE-150增殖、衰老和凋亡的影响,并研究相关机制。方法将KYSE-150细胞与CFZ共孵育24 h,然后采用CCK-8法评估CFZ的毒性作用并筛选研究浓度。分别使用CFZ、^(125)I粒子照射(6 Gy),以及CFZ和^(125)I粒子的联合治疗处理KYSE-150细胞。通过EdU法检测细胞增殖,β-半乳糖苷酶染色检测细胞衰老,流式细胞术检测细胞周期和凋亡。通过蛋白免疫印迹法检测与DNA损伤、内质网应激、凋亡和凋亡通路相关的蛋白表达。通过siRNA沉默CHOP基因后,细胞经联合治疗处理,随后评估凋亡相关蛋白的改变。结果CFZ浓度≥20 nmol/L时,细胞活力受限。20 nmol/L的CFZ处理24 h对细胞增殖、DNA损伤、细胞衰老和凋亡的影响不显著。相较于^(125)I粒子单一治疗,联合治疗进一步抑制细胞增殖,加重DNA损伤,下调S期和G2/M期细胞比例,加剧内质网应激,促进细胞凋亡。蛋白水平研究显示,CFZ通过内源性凋亡通路促进^(125)I粒子诱导的细胞凋亡。联合治疗诱导的内源性细胞凋亡由PERK-eIF2α-ATF4-CHOP通路介导,且不依赖于p53。此外,CFZ能促使^(125)I粒子诱导的衰老细胞死亡。结论CFZ联合^(125)I粒子照射能抑制细胞增殖、加重DNA损伤和内质网应激、通过PERK-eIF2α-ATF4-CHOP通路促进细胞凋亡、并促使衰老细胞死亡;CFZ是^(125)I粒子有效的增敏剂。Objective To investigate the effects of carfilzomib(CFZ)combined with iodine-125(^(125)I)seed irradiation on the proliferation,cellular senescence,and apoptosis in human esophageal cancer cells KYSE-150,and study the related mechanisms.Methods KYSE-150 cells were incubated with CFZ for 24 h,and then CCK-8 assay was used to evaluate the toxic effect of CFZ and screen the research concentration.KYSE-150 cells were treated with CFZ,^(125)I seed irradiation(6 Gy),and combined treatment with CFZ and^(125)I seeds,respectively.Cell proliferation was detected by EdU assay,cellular senescence was detected byβ-galactosidase staining,and cell cycle and apoptosis were determined by flow cytometry.The expression of proteins related to DNA damage,endoplasmic reticulum stress,apoptosis,and apoptosis pathways was evaluated by Western blotting.After silencing the CHOP gene by siRNA,cells were treated with combined treatment,and then changes in apoptosis-related proteins were evaluated.Results Cell viability was limited when the concentration of CFZ was≥20 nmol/L.Treatment with 20 nmol/L CFZ for 24 h had no significant effect on cell proliferation,DNA damage,cellular senescence,and apoptosis.Compared with^(125)I seeds monotherapy,combined treatment further inhibited cell proliferation,aggravated DNA damage,downregulated the proportion of cells in the S phase and G2/M phase,aggravated endoplasmic reticulum stress,and promoted cell apoptosis.Experiments at the protein level showed that CFZ promoted^(125)I seeds-induced apoptosis through the intrinsic apoptotic pathway.The intrinsic apoptosis induced by the combined treatment was mediated by the PERK-eIF2α-ATF4-CHOP pathway and was independent of p53.In addition,CFZ promoted the death of^(125)I seeds-induced senescent cells.Conclusion CFZ combined with^(125)I seed irradiation can inhibit cell proliferation,aggravate DNA damage and endoplasmic reticulum stress,promote cell apoptosis through the PERK-eIF2α-ATF4-CHOP pathway,and promote the death of senescent cells.CFZ is an effect

关 键 词：碘-125粒子 卡非佐米 食管癌 凋亡 细胞衰老 

分 类 号：R735.1[医药卫生—肿瘤]