电离辐射诱导人肠上皮细胞焦亡机制  

Mechanisms of ionizing radiation⁃induced pyroptosis in human intestinal epithelial cells

作　　者：王琼 洪剑[3] 郭瑾[4] 段君昭 丁可昕 台福敏 郑晓飞 葛常辉 WANG Qiong;HONG Jian;GUO Jin;DUAN Junzhao;DING Kexin;TAI Fumin;ZHENG Xiaofei;GE Changhui(School of Basic Medical Sciences,Anhui Medical University,Hefei 230032,China;Beijing Key Laboratory for Radiobiology,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China;the Eighth Medical Center,Chinese PLA General Hospital,Beijing 100091,China;Southern Medical District,Chinese PLA General Hospital,Beijing 100071,China)

机构地区：[1]安徽医科大学基础医学院,合肥230032 [2]军事科学院军事医学研究院放射生物学北京市重点实验室,北京100850 [3]解放军总医院第八医学中心,北京100091 [4]解放军总医院京南医疗区,北京100071

出　　处：《军事医学》2024年第4期241-250,共10页Military Medical Sciences

摘　　要：目的探讨消皮素E(GSDME)介导的细胞焦亡参与辐射诱导的肠损伤分子机制及消皮素(GSDM)蛋白家族是否通过通用的信号通路调控细胞焦亡。方法利用人正常结肠上皮NCM460细胞和人结肠癌HT-29细胞辐射不同剂量及辐射后不同时间,通过观察焦亡小泡、细胞存活、焦亡执行蛋白的切割进行焦亡指标的检测,对HT-29细胞过表达GSDME并辐射后通过RNA测序技术,对焦亡相关差异基因进行富集分析,筛选相关通路差异基因进行验证。结果辐射诱导NCM460细胞发生显著的细胞焦亡,HT-29细胞过表达GSDME后辐射诱导GSDME激活发生显著的细胞焦亡。成功模拟人肠细胞焦亡状态,过表达GSDME-N和GSDMD-N具有超过50%的焦亡状态下的差异基因;测序分析显示,焦亡状态下的基因主要富集在免疫反应、炎症反应和Rap1信号通路等。结论GSDME激活介导了辐射诱导肠细胞焦亡的发生,GSDM蛋白家族通过通用的调控模式参与细胞焦亡,辐射通过免疫反应、炎症反应和Rap1信号通路诱导GSDME/D激活调控细胞焦亡。Objective To investigate the mechanism underlying gasdermin E(GSDME)-mediated pyroptosis in radiation-induced intestinal injury and to find out whether gasdermin(GSDM)family members regulate pyroptosis through similar signaling pathways.Methods Human normal colon epithelial cells(NCM460)and human colon cancer cells(HT-29)were exposed to radiation of different doses and durations before pyroptosis indicators were evaluated by observing pyroptotic bubbles,cell survival,and the cleavage of pyroptosis execution proteins.HT-29 cells overexpressing GSDME were subjected to radiation,followed by enrichment analysis of pyroptosis-related differentially expressed genes using RNA-seq.Results Radiation induced substantial pyroptosis in NCM460 cells.Overexpression of GSDME in HT-29 cells resulted in substantial radiation-induced pyroptosis.The pyroptosis state of human intestinal cells was simulated in the HT-29 model cell line.Overexpressions of GSDME-N and GSDMD-N resulted in the expression of more than 50% of the differentially expressed genes in the pyroptosis state.Sequencing analysis showed that the genes in the pyroptosis state were mainly overrepresented in immune response,inflammatory response,and Rap1 signaling pathway.Conclusion GSDME activation can mediate radiation-induced pyroptosis by producing GSDME-N fragments.GSDM family members participate in pyroptosis in a similar mode of regulation.Furthermore,radiation-induced activation of GSDME/D may regulate pyroptosis through immune response,inflammatory response,and Rap1 signaling pathway.

关 键 词：细胞焦亡 电离辐射 RNA测序 消皮素 肠细胞 

分 类 号：R818.74[医药卫生—放射医学]