去泛素化酶头帕肿瘤综合征蛋白对肺癌预后及肺泡巨噬细胞极化的影响  

Effect of depubiquitinase cylindromatosis on the prognosis of lung cancer and the polarization of alveolar macrophages

作　　者：张媛媛[1] 朱柠[1] ZHANG Yuanyuan;ZHU Ning(Department of Respiratory Diseases,Huashan Hospital Affiliated to Fudan University,Shanghai 200040,China)

机构地区：[1]复旦大学附属华山医院呼吸科,上海200040

出　　处：《肿瘤基础与临床》2020年第1期7-12,共6页journal of basic and clinical oncology

基　　金：上海市卫生和计划生育委员会科研课题青年项目(20164Y0139)。

摘　　要：目的探讨去泛素化酶头帕肿瘤综合征蛋白(CYLD)对肺癌临床预后及肺泡巨噬细胞极化的影响,并探索其可能分子机制。方法首先,利用Kaplan-Meier Plotter在线数据库分析CYLD表达与肺癌生存时间的关系。其次,体外培养大鼠肺泡巨噬细胞NR8383,设计小干扰RNA侵染肺泡巨噬细胞,敲低CYLD的表达,根据干预条件不同分为4组sicontrol组;sicontrol+罗氟司特组;siCYLD组;siCYLD+罗氟司特组。每组均予以10 mg/L脂多糖处理。采用荧光定量PCR方法检测M1、M2型肺泡巨噬细胞相关炎症因子白介素-12(IL-12)、肿瘤坏死因子-α(TNF-α),IL-8、转化生长因子-β(TGF-β)mRNA的表达水平;采用Western blot方法检测核转录因子-κB(NF-κB)的磷酸化水平。结果CYLD表达与肺癌生存时间呈明显正相关,亚组分析显示肺腺癌患者获益更明显(P<0.05)。相较于sicontrol组,敲低CYLD后,M1型肺泡巨噬细胞相关炎症因子IL-12、TNF-α表达增加(P<0.05),M2型肺泡巨噬细胞相关炎症因子IL-8、TGF-β表达下降(P<0.05);NF-κB的磷酸化水平增高(P<0.05);而罗氟司特可部分逆转此过程(P<0.05)。结论CYLD与肺癌患者总体生存时间关系密切,且可通过间接抑制NF-κB的磷酸化水平,抑制肺泡巨噬细胞向M1型极化,诱导其向M2型极化。Objective To investigate the effect of cylindromatosis(CYLD)on the survival time of lung cancer and the polarization of alveolar macrophages,and to explore the possible molecular mechanism.Methods Firstly,Kaplan-Meier Plotter online database was used to analyze the effect of CYLD on survival time of lung cancer.Secondly,the rat alveolar macrophages NR8383 were cultured in vitro.Small interfere RNA were designed to infect the alveolar macrophages and knock down the expression of CYLD.According to the different intervention conditions,they were divided into four groups:the sicontrol group;the sicontrol+roflumilast group;the siCYLD group;the siCYLD+roflumilast group.Each group was treated with 10 mg/L lipopolysaccharide.Quantitative PCR was used to detect the mRNA expression levels of M1,M2 related inflammatory factors[interleukin-12(IL-12),tumor necrosis factor-α(TNF-α),IL-8;transforming growth factor-β(TGF-β)]in alveolar macrophages,Western blot was used to detect the phosphorylation level of nuclear factorκB(NF-κB).Results The expression of CYLD mRNA was positively correlated with the survival time of lung cancer.Subgroup analysis showed that patients with lung adenocarcinoma benefited more significantly(P<0.05).After knocking down the CYLD expression,the mRNA expression of IL-12,TNF-αin the M1 type alveolar macrophages increased(P<0.05),but the mRNA expression of IL-8,TGF-βin the M2 type alveolar macrophages decreased(P<0.05).Meanwhile,the phosphorylation level of NF-κB increased(P<0.05).These changes could be partially reversed by roflumilast(P<0.05).Conclusion CYLD is significantly related to the overall survival time of lung cancer patients.CYLD could inhibit the polarization of alveolar macrophages to M1,induce them to M2,and decrease the inflammatory level of alveolar microenvironment by indirectly inhibiting the phosphorylation level of NF-κB.

关 键 词：去泛素化酶 头帕肿瘤综合征蛋白 肺泡巨噬细胞 核转录因子-κB 

分 类 号：R734.2[医药卫生—肿瘤]