低氧条件下SIRT3调控NF-κB通路抑制肺癌细胞转移和炎症反应  

SIRT3 regulates the NF-κB pathway inhibiting lung cancer cell metastasis and inflammatory response under hypoxia

作　　者：黄波[1,2] 丁洁 郭红荣[1] 王红娟 徐建群[1] 郑泉 HUANG Bo;DING Jie;GUO Hongrong;WANG Hongjuan;XU Jianqun;ZHENG Quan(Wuhan Third Hospital/Tongren Hospital of Wuhan University,Hubei Wuhan 430074,China;Wuhan East Lake High-Tech Development Zone Jiufeng Street Center City Community Health Service Center,Hubei Wuhan 430075,China)

机构地区：[1]武汉市第三医院/武汉大学附属同仁医院,湖北武汉430074 [2]武汉东湖新技术开发区九峰街中心城社区卫生服务中心,湖北武汉430075

出　　处：《现代肿瘤医学》2023年第24期4508-4514,共7页Journal of Modern Oncology

基　　金：湖北省卫生健康委员会科研项目基金(编号:WJ2021F005);湖北省武汉市卫生健康委员会医学科研项目基金(编号:WX21Q43);武汉中青年医学骨干人才培养工程(编号:武卫通[2019]87号)。

摘　　要：目的:探讨低氧条件下肺癌细胞中沉默信息调节因子3(SIRT3)过表达调控核因子κB(NF-κB)通路对细胞炎症反应、迁移和侵袭的影响。方法:基于A549肺癌细胞,根据SIRT3的序列构建过表达和空载慢病毒转染肺癌细胞,未转染的细胞为对照,暴露于常氧(21%O_(2))和低氧(5%~8%O_(2))条件下培养,验证实验可行。根据处理方法将细胞分为常氧组、低氧组、JSH-23(NF-κB转录活性抑制剂)组、SIRT3过表达组、JSH-23+SIRT3过表达组。MTT法检测细胞增殖,Transwell法检测细胞侵袭,细胞划痕法检测细胞迁移,Western-blot检测蛋白MMP-2、ICAM-1、SIRT3和p-NF-κB p65的表达水平,qPCR检测细胞中SIRT3、p-NF-κB p65的mRNA表达,ELISA检测细胞中IL-1β、TNF-α和IL-6含量。结果:与常氧组相比,低氧组中细胞增殖能力增强(P<0.05),侵袭和迁移能力上升(P<0.05),MMP-2、ICAM-1和p-NF-κB p65蛋白表达上调(P<0.05),p-NF-κB p65的mRNA表达上调(P<0.05),SIRT3 mRNA和蛋白表达下调(P<0.05),IL-1β、TNF-α和IL-6含量上升(P<0.05)。与低氧组相比,JSH-23组和SIRT3过表达组细胞增殖能力、侵袭和迁移能力下降(P<0.05),MMP-2、ICAM-1和p-NF-κB p65蛋白表达下调(P<0.05),p-NF-κB p65的mRNA表达下调(P<0.05),SIRT3 mRNA和蛋白表达上调(P<0.05),IL-1β、TNF-α和IL-6含量下降(P<0.05)。与JSH-23组相比,JSH-23+SIRT3过表达组变化趋势更加显著(P<0.05)。结论:低氧条件下SIRT3抑制NF-κB通路,从而抑制肺癌细胞的炎症反应和进展。Objective:To investigate the effect of silent information regulator 3(SIRT3)overexpression regulating nuclear factor-κB(NF-κB)pathway on cellular inflammatory response,migration and invasion in lung cancer cells under hypoxic conditions.Methods:Based on A549 lung cancer cells,overexpressed and null lentiviral transfected lung cancer cells were constructed according to the sequence of SIRT3,and untransfected cells were used as controls and exposed to normoxic(21%O_(2))and hypoxic(5%~8%O_(2))conditions in culture to verify the feasibility of the experiment.The cells were divided into normoxia,hypoxia,JSH-23(NF-κB transcriptional activity inhibitor),SIRT3 overexpression,and JSH-23+SIRT3 overexpression groups according to the treatments.cell proliferation was detected by MTT,cell invasion by Transwell,cell migration by cell scratching,and protein MMP-2,ICAM-1,SIRT3 and p-NF-κB p65 by Western-blot.qPCR was performed to detect the mRNA expression of SIRT3 and p-NF-κB p65 in the cells,and ELISA was performed to detect the content of IL-1β,TNF-αand IL-6 in the cells.Results:Compared with the normoxia group,cell proliferation ability increased(P<0.05),invasion and migration ability increased(P<0.05),MMP-2,ICAM-1 and p-NF-κB p65 protein expression was upregulated(P<0.05),mRNA expression of p-NF-κB p65 was upregulated(P<0.05),SIRT3 mRNA and protein expression was downregulated(P<0.05),and IL-1β,TNF-αand IL-6 levels were increased(P<0.05).Compared with the hypoxic group,the JSH-23 group and SIRT3 overexpression group showed decreased cell proliferation,invasion and migration ability(P<0.05),down-regulation of MMP-2,ICAM-1 and p-NF-κB p65 protein expression(P<0.05),down-regulation of p-NF-κB p65 mRNA expression(P<0.05),and SIRT3 mRNA and protein expression was upregulated(P<0.05),and IL-1β,TNF-αand IL-6 levels were decreased(P<0.05).The trend of change was more significant in the JSH-23+SIRT3 overexpression group compared with the JSH-23 group(P<0.05).Conclusion:SIRT3 inhibits the NF-κB pathway under hypoxic c

关 键 词：肺癌 低氧环境 SIRT3过表达 NF-ΚB通路 炎症反应 

分 类 号：R73-3[医药卫生—肿瘤]