重组新城疫病毒rL-RVG抑制Nrf2-GCLC-GPX4通路诱导胃癌细胞铁死亡  

Recombinant Newcastle disease virus rL-RVG induces ferroptosis of gastric cancer cells through inhibiting Nrf2-GCLC-GPX4 pathway

作　　者：贡克文 田仪督 何瑛珏 李洋 步雪峰[1] GONG Kewen;TIAN Yidu;HE Yingjue;LI Yang;BU Xuefeng(Department of General Surgery,Affiliated People’s Hospital of Jiangsu University,Zhenjiang,Jiangsu Province,212000,China;Department of Respiratory Medicine,Affiliated People’s Hospital of Jiangsu University,Zhenjiang,Jiangsu Province,212000,China)

机构地区：[1]江苏大学附属人民医院普外科,江苏镇江212000 [2]江苏大学附属人民医院呼吸内科,江苏镇江212000

出　　处：《陆军军医大学学报》2024年第13期1485-1493,共9页Journal of Army Medical University

基　　金：镇江市科技创新资金项目(SH2023004)。

摘　　要：目的探讨重组新城疫病毒(recombinant Newcastle disease virus,rL-RVG)是否通过Nrf2-GCLC-GPX4通路引发胃癌细胞铁死亡。方法培养人胃癌HGC-27细胞,分别用rL-RVG、新城疫病毒(Newcastle disease virus,NDV)、PBS溶液(对照组)处理HGC-27细胞后,用CCK-8、Transwell侵袭、细胞划痕实验检测胃癌HGC-27细胞的增殖、侵袭、迁移能力。加入铁死亡促进剂(erastin)、核因子E2相关因子2(nuclear factor E2 related factor 2,Nrf2)促进剂(TBHQ)及抑制剂(ML385)作为干预组,脂质氧化试剂盒检测各组丙二醛(malondialdehyde,MDA)含量;DCFH-DA荧光探针及流式细胞仪检测活性氧(reactive oxygen species,ROS)的含量;Western blot、免疫荧光法检测Nrf2-GCLC-GPX4通路相关蛋白表达情况。结果与对照组比较,rL-RVG和NDV处理后,细胞的存活率均下降,呈剂量、时间依赖性,且rL-RVG处理组较为显著(P<0.05);NDV和rL-RVG处理组胃癌HGC-27细胞迁移、侵袭能力均受到抑制,后者较前者抑制更明显(P<0.05);与对照组比较,病毒组Nrf2、GCLC、SLC7A11、GPX4蛋白表达下降(P<0.05),MDA水平、ROS相对表达量上升(P<0.05),rL-RVG组上升或下降的趋势较NDV组明显,且erastin组SLC7A11、GPX4蛋白表达下降(P<0.05);与对照组比较,TBHQ组、ML385组Nrf2、GCLC、SLC7A11、GPX4蛋白表达分别上升、下降(P<0.05),MDA水平、ROS相对表达量分别下降、上升(P<0.05);与rL-RVG组比较,rL-RVG+TBHQ组、rL-RVG+ML385组Nrf2、GCLC、SLC7A11、GPX4蛋白表达分别上升、下降(P<0.05),MDA水平、ROS相对表达量分别下降、上升(P<0.05)。结论重组新城疫病毒(rL-RVG)可通过Nrf2-GCLC-GPX4通路诱导胃癌细胞铁死亡的发生,抑制肿瘤细胞的生长。Objective To investigate whether recombinant Newcastle disease virus(rL-RVG)induces iron death in gastric cancer cells through Nrf2-GCLC-GPX4 pathway.Methods After human gastric cancer HGC-27 cells were treated with rL-RVG,Newcastle disease virus(NDV)and PBS solution(control group),respectively,cell proliferation,invasion and migration were detected by CCK-8 assay and Transwell invasion assay and cell scratch test.Ferroptosis accelerator(erastin),and nuclear factor E2 related factor 2(Nrf2)accelerator(TBHQ)and inhibitor(ML385)were added respectively as controls.The content of malondialdehyde(MDA)in each treatment group was detected by lipid oxidation kit.The content of reactive oxygen species(ROS)was detected by DCFH-DA fluorescent probe and flow cytometry.Western blotting and immunofluorescence assay were employed to measure the expression levels of Nrf2-GCLC-GPX4 pathway related proteins.Results Compared with the control group,the survival rate of HGC-27 cells were significantly decreased after rL-RVG and NDV treatment in a dose-and time-dependent manner,and the effect was more significant in the rL-RVG treatment group(P<0.05).The migration and invasion abilities of HGC-27 cells were obviously inhibited in the NDV and rL-RVG treatment groups,and the latter had more notable inhibition than the former.The protein levels of Nrf2,GCLC,SLC7A11 and GPX4 were statistically decreased(P<0.05),and the contents of MDA and ROS were increased(P<0.05)in the virus treatment groups than the control group,with the increasing or decreasing trend more significant in the rL-RVG group than the NDV group.What’s more,the protein levels of SLC7A11 and GPX4 were decreased in the erastin group(P<0.05).Compared with the control group,those of Nrf2,GCLC,SLC7A11 and GPX4 were increased in the TBHQ group and decreased in the ML385 group(P<0.05),while the contents of MDA and ROS were decreased and increased respectively in the above 2 groups(P<0.05).Compared with the rL-RVG group,the rL-RVG+TBHQ group and rL-RVG+ML385 group had enhanced an

关 键 词：重组新城疫病毒 胃癌细胞 铁死亡 核因子E2相关因子2 谷胱甘肽过氧化物酶4 

分 类 号：R394.3[医药卫生—医学遗传学] R73-362[医药卫生—基础医学] R735.2