Article(id=1208106713203445846, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1208106710208717234, articleNumber=null, orderNo=null, doi=10.11855/j.issn.0577-7402.2022.04.0407, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1617120000000, receivedDateStr=2021-03-31, revisedDate=null, revisedDateStr=null, acceptedDate=1623686400000, acceptedDateStr=2021-06-15, onlineDate=1765964685964, onlineDateStr=2025-12-17, pubDate=1651075200000, pubDateStr=2022-04-28, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1765964685964, onlineIssueDateStr=2025-12-17, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1765964685964, creator=13701087609, updateTime=1765964685964, updator=13701087609, issue=Issue{id=1208106710208717234, tenantId=1146029695717560320, journalId=1189873630562394117, year='2022', volume='47', issue='4', pageStart='321', pageEnd='426', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1765964685250, creator=13701087609, updateTime=1765964685250, updator=13701087609, preIssue=null, nextIssue=null, ext=null, issueFiles=null}, startPage=407, endPage=411, ext={EN=ArticleExt(id=1208106713450909799, articleId=1208106713203445846, tenantId=1146029695717560320, journalId=1189873630562394117, language=EN, title=Research progress on the role of neutrophil extracellular traps in tumor microenvironment, columnId=1190243275882729994, journalTitle=Medical Journal of Chinese People’s Liberation Army, columnName=Review, runingTitle=null, highlight=null, articleAbstract=

Neutrophil extracellular trap (NETs) is a kind of DNA-based skeleton released by neutrophils to the outside of the cell, in which a variety of granular proteins, proteolytic enzymes, antibacterial peptides, and other network structures are embedded.In recent years, studies have confirmed that NETs are highly expressed in a variety of tumors, and tumor microenvironment (TME)is a special environment for tumor cell survival and development, including cellular and non-cellular components. This article mainly explains that TME can stimulate the formation of NETs, and NETs can also promote tumor proliferation, invasion and metastasis by affecting tumor cells, immune cells, endothelial cells, epithelial cells, fibroblasts and extracellular matrix in TME. Finally, the clinical application prospects of NETs as tumor markers and potential therapeutic targets are discussed.

, correspAuthors=De-Kui Zhang, authorNote=null, correspAuthorsNote=
*E-mail:
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中性粒细胞外诱捕网(NETs)是中性粒细胞受到各种刺激后产生的以DNA为骨架,其间镶嵌各种颗粒蛋白、蛋白水解酶及抗菌肽等的网状结构。近年来的研究证实,NETs在多种肿瘤中高表达,而肿瘤微环境(TME)是肿瘤细胞生存和发展的特殊环境,包括细胞成分及非细胞成分。TME中的白细胞介素(IL)-8及肿瘤相关成纤维细胞产生的淀粉样蛋白β能刺激NETs的形成,而NETs同时通过影响TME中肿瘤细胞的生物学特性、免疫细胞杀伤肿瘤的能力、内皮细胞的表型、肿瘤相关成纤维细胞的功能及细胞外基质的组分等促进肿瘤增殖、侵袭及转移。本文从以上几个方面对NETs在TME中的作用进行综述,以探讨NETs作为肿瘤标志物及潜在治疗靶点的临床应用前景。

, correspAuthors=张德奎, authorNote=null, correspAuthorsNote=
张德奎,E-mail:
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马雪妮,博士研究生,主要从事中性粒细胞外诱捕网与消化系统疾病的关系研究

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马雪妮,博士研究生,主要从事中性粒细胞外诱捕网与消化系统疾病的关系研究

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马雪妮,博士研究生,主要从事中性粒细胞外诱捕网与消化系统疾病的关系研究

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中性粒细胞外诱捕网在肿瘤微环境中的作用研究进展
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马雪妮 , 许慧梅 , 张德奎 *
解放军医学杂志 | 综述 2022,47(4): 407-411
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解放军医学杂志 | 综述 2022, 47(4): 407-411
中性粒细胞外诱捕网在肿瘤微环境中的作用研究进展
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马雪妮, 许慧梅, 张德奎*
作者信息
  • 兰州大学第二医院消化内科,兰州 730030
  • 马雪妮,博士研究生,主要从事中性粒细胞外诱捕网与消化系统疾病的关系研究

通讯作者:

张德奎,E-mail:
Research progress on the role of neutrophil extracellular traps in tumor microenvironment
Xue-Ni Ma, Hui-Mei Xu, De-Kui Zhang*
Affiliations
  • Department of Gastroenterology, the Second Hospital of Lanzhou University, Lanzhou 730030, China
出版时间: 2022-04-28 doi: 10.11855/j.issn.0577-7402.2022.04.0407
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中性粒细胞外诱捕网(NETs)是中性粒细胞受到各种刺激后产生的以DNA为骨架,其间镶嵌各种颗粒蛋白、蛋白水解酶及抗菌肽等的网状结构。近年来的研究证实,NETs在多种肿瘤中高表达,而肿瘤微环境(TME)是肿瘤细胞生存和发展的特殊环境,包括细胞成分及非细胞成分。TME中的白细胞介素(IL)-8及肿瘤相关成纤维细胞产生的淀粉样蛋白β能刺激NETs的形成,而NETs同时通过影响TME中肿瘤细胞的生物学特性、免疫细胞杀伤肿瘤的能力、内皮细胞的表型、肿瘤相关成纤维细胞的功能及细胞外基质的组分等促进肿瘤增殖、侵袭及转移。本文从以上几个方面对NETs在TME中的作用进行综述,以探讨NETs作为肿瘤标志物及潜在治疗靶点的临床应用前景。

中性粒细胞外诱捕网  /  肿瘤微环境  /  研究进展

Neutrophil extracellular trap (NETs) is a kind of DNA-based skeleton released by neutrophils to the outside of the cell, in which a variety of granular proteins, proteolytic enzymes, antibacterial peptides, and other network structures are embedded.In recent years, studies have confirmed that NETs are highly expressed in a variety of tumors, and tumor microenvironment (TME)is a special environment for tumor cell survival and development, including cellular and non-cellular components. This article mainly explains that TME can stimulate the formation of NETs, and NETs can also promote tumor proliferation, invasion and metastasis by affecting tumor cells, immune cells, endothelial cells, epithelial cells, fibroblasts and extracellular matrix in TME. Finally, the clinical application prospects of NETs as tumor markers and potential therapeutic targets are discussed.

neutrophil extracellular trap  /  tumor microenvironment  /  research progress
马雪妮, 许慧梅, 张德奎. 中性粒细胞外诱捕网在肿瘤微环境中的作用研究进展. 解放军医学杂志, 2022 , 47 (4) : 407 -411 . DOI: 10.11855/j.issn.0577-7402.2022.04.0407
Xue-Ni Ma, Hui-Mei Xu, De-Kui Zhang. Research progress on the role of neutrophil extracellular traps in tumor microenvironment[J]. Medical Journal of Chinese People’s Liberation Army, 2022 , 47 (4) : 407 -411 . DOI: 10.11855/j.issn.0577-7402.2022.04.0407
肿瘤微环境(tumor microenvironment,TME)是肿瘤生长所必需的微环境[1],包括细胞成分与非细胞成分。细胞成分包括间充质细胞、免疫细胞、成纤维细胞、内皮细胞、周细胞及脂肪细胞等,非细胞成分包括细胞外基质、基质金属蛋白酶(MMPs)、细胞因子及趋化因子等。其中免疫细胞、间充质细胞及内皮细胞可分泌多种细胞因子直接作用于肿瘤细胞,如肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)及转化生长因子-β(transforming growth factor-β,TGF-β)等,它们相互调控,共同形成抑制性免疫分子网络,使肿瘤细胞能够躲避机体免疫系统的攻击,实现免疫逃逸[2-3]
中性粒细胞是人外周血中最丰富的白细胞,约占白细胞总数的50%~70%,在先天性免疫系统中扮演着重要角色[4],可通过吞噬作用、脱颗粒、产生活性氧等多种途径抵御外界病原体的入侵,是机体应对病原体入侵的第一道防线[5]。除此以外,中性粒细胞还可形成有别于细胞坏死及凋亡的特殊细胞死亡程序——中性粒细胞外诱捕网(neutrophil extracellular traps,NETs),参与机体的各种病理生理过程[6]
近年来研究发现,中性粒细胞产生及释放NETs的过程不仅发生在败血症等感染性疾病中,而且在非感染性疾病如系统性红斑狼疮(SLE)、类风湿关节炎(RA)、糖尿病、动脉粥样硬化、急性肺损伤、肝缺血再灌注损伤、血栓形成及慢性肝病中也发挥着关键作用[7-8]。此外,NETs在多种肿瘤中高表达,如肺癌[9-11]、肝癌[11]、淋巴瘤[12]、结直肠癌[13]、乳腺癌[10,14]、胃癌[15]等,并与肿瘤的发生、发展、转移及其相关的血栓形成密切相关。本文就TME对NETs表达的影响,以及NETs在TME中的作用进行综述,旨在探讨NETs作为肿瘤标志物及潜在治疗靶点的临床应用前景。
1996年,Takei等[6]在使用佛波酯(phorbol myristate acetate,PMA)刺激中性粒细胞时,观察到一种不同于细胞凋亡(核固缩、细胞质空泡化)及细胞坏死(保持核膜完整性)的现象,即中性粒细胞的分叶核退缩、扩散,继而核膜破裂,核内成分释放到细胞质,细胞膜破裂[6]。直到2004年,Brinkmann等[16]将该过程中释放到胞外的纤维网状结构正式命名为NETs。NETs是以DNA为骨架并镶嵌多种活性蛋白的细胞外网状结构,其中的活性蛋白包括中性粒细胞弹性蛋白酶(neutrophil elastase,NE)、基质金属蛋白酶9(matrix metalloprotease-9,MMP-9)、髓过氧化物酶(myeloperoxidase,MPO)及组织蛋白酶G(cathepsin G,CG)等,而染色质是网状结构的主干[17]。研究发现,多种物质均可刺激中性粒细胞产生NETs,包括细菌、真菌、原生动物、病毒等在内的病原体以及一些化学或生物因素[5,9-13]
白细胞介素(IL)-8是CXC家族的典型趋化因子,又称CX-CL8,负责中性粒细胞向炎症部位的募集并活化[18]。研究发现,来自弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)的IL-8与中性粒细胞上的受体(CXCR2)相互作用,通过激活Src、p38及ERK信号通路形成NETs[12]。同时,Alfaro等[19]发现,在前列腺癌及非小细胞肺癌中,肿瘤细胞产生的包括IL-8在内的趋化因子能够募集骨髓来源的抑制性细胞(myeloid-derived suppressor cells,MDSCs),使其形成NETs,该过程能够募集更多的中性粒细胞,有助于肿瘤细胞的免疫逃避。最近有研究显示,黑色素瘤及胰腺癌中的肿瘤相关成纤维细胞(cancer-associated fibroblasts,CAFs)产生的淀粉样蛋白β能够刺激NETs的形成,称为肿瘤相关NETs(t-NETs),此过程依赖于活性氧(ROS)的产生[20]。因此,TME能够刺激NETs的产生,从而促进NETs形成(NETosis)。
NETs能够促进肿瘤细胞的增殖。Miller-Ocuin等[21]研究发现,来源于NETs的细胞外DNA可通过激活胰腺星状细胞促进肿瘤细胞增殖,而通过肽酰基精氨酸脱亚胺酶4(peptidylarginine deiminase 4,PAD4)抑制剂或脱氧核糖核酸酶Ⅰ(DNase Ⅰ)抑制NETs的生成能够逆转上述作用;且这种促肿瘤细胞增殖的机制还可能涉及NETs的多种活性蛋白酶。Yazdani等[22]研究发现,结直肠癌细胞受到外界刺激后会释放损伤相关分子模式(DAMP)蛋白,在此过程中将中性粒细胞募集到TME,并诱导NETs的形成,而NETs主要通过增加线粒体的生物合成来增加能量的产生,并促进肿瘤细胞的增殖。NETs通过释放NE激活TLR4-p38-PGC-1α轴,从而促进线粒体生物合成及肿瘤生长。在PAD4-/-小鼠体内,通过DNase Ⅰ或NE抑制剂(NEi)抑制NETs的形成,或在体外阻断NE-TLR4-PGC-1α轴,都可以抑制线粒体的生物合成并减缓肿瘤的生长。此外,DLBCL产生的NETs可直接上调DLBCL Toll样受体9(Toll-like receptor 9,TLR9)途径,随后激活核因子(NF)-кB、STAT3及p38途径,从而促进肿瘤进展。动物实验研究发现,抑制NETs形成、阻断IL8-CXCR2轴或抑制TLR9均会抑制肿瘤的进展[12]
NETs可促进肿瘤细胞的转移。Martins-Cardoso等[14]发现,NETs能够将人乳腺癌细胞MCF7(主要为CD44Lo/–CD24+表型)转化为CD44+CD24Lo/–表型,而CD44是一种细胞表面糖蛋白受体,可识别多种配体,包括细胞外基质成分如透明质酸、骨桥蛋白、金属蛋白酶等,与肿瘤迁移及转移有关[23]。将NETs与人乳腺癌细胞MCF7共培养发现,NETs能将MCF7典型的上皮形态转变为间充质表型,即N-钙黏蛋白及纤连蛋白的表达增加,而E-钙黏蛋白的表达被抑制,从而导致MCF7细胞的迁移能力增强。还有研究证实,在小鼠结肠癌及肺癌模型中,即便不存在败血症,NETs仍可捕获循环中的肿瘤细胞,使其黏附于肝脏[24]。而Najmeh等[25]研究发现,在肺腺癌细胞中,这种黏附作用是整合素介导的蛋白-蛋白互相作用;在NETs及肿瘤细胞中,β1整合素均呈高表达,且两种β1整合素均参与了这种黏附作用;而将NETs上的β1整合素阻断后,肿瘤细胞与NETs黏附减少,当肿瘤来源的β1整合素被抑制后,来源于NETs的β1整合素也会相应减少,由此可以推论来源于NETs与来源于肿瘤细胞的β1整合素可能存在某种共域化作用。此外,跨膜蛋白CCDC25是一种存在于肿瘤细胞表面的NETs DNA受体,可通过激活ILK-β-parvin信号通路增强肿瘤细胞的活性,从而促进肿瘤的转移[26]
TME中浸润的免疫抑制性细胞是肿瘤抑制性免疫微环境形成的关键因素,这些免疫抑制性细胞包括肿瘤相关巨噬细胞(tumor associated macrophages,TAMs)、MDSCs、调节性T细胞(regulatory T cells,Tregs)等,它们能够协助肿瘤进行免疫逃逸,促进肿瘤的发生发展[27]。研究发现,肿瘤细胞产生的趋化因子可促进中性粒细胞及MDSCs发生NETosis,在此过程中,NETs将肿瘤细胞包裹起来,阻断了免疫细胞与周围靶细胞的结合,从而使肿瘤细胞免受CD8+ T细胞及自然杀伤(NK)细胞介导的细胞毒性作用,而PAD4抑制剂可抑制肿瘤的转移,是肿瘤免疫治疗的基础[13]。Xia等[28]发现,表达DNase Ⅰ的腺病毒(即AAV-DNaseⅠ)可通过抑制中性粒细胞募集、抑制NETs形成、增加CD8+ T细胞数量、减少CD4+ T细胞数量来调节TME中的先天性及获得性免疫应答,而CD8+ T细胞是抗原特异性抗肿瘤免疫的主要效应器,可以识别并杀死肿瘤细胞。此外,有研究发现,肿瘤细胞通过募集中性粒细胞使其分泌CCL17而将Tregs募集到肿瘤部位,此过程损害了免疫系统对肿瘤的杀伤能力[29]。因此,NETs可能通过作用于TME中的免疫细胞协助肿瘤进行免疫逃逸,从而促进肿瘤的发生发展。
NETs中的多种成分可通过影响内皮细胞而促进肿瘤微血管的生成。Jung等[30]发现,组蛋白作为NETs的重要组成部分,能在胰腺癌中促进内皮细胞小管形成,而组蛋白的抑制剂(如肝素或多聚唾液酸)可逆转以上作用。此外,NE能通过诱导TME内的血管生成促进肿瘤细胞的增殖、迁移及侵袭,其可能的机制是NE导致肿瘤抑制因子失活,从而促进TME中的血管生成[31]。Wilson等[32]认为CG不仅具有降解细胞外基质的功能,还可通过刺激血管内皮生长因子(vascular endothelial growth factor,VEGF)而促进新生血管形成,使转移灶拥有丰富的血供,从而继续生长;对骨髓转移癌小鼠模型使用CG抑制剂,可使其微血管密度低于正常对照。另外,有研究显示,结直肠癌患者肿瘤细胞产生的NETs能够促进人脐静脉内皮细胞转化为促凝表型,即表现为磷脂酰丝氨酸(PS)外露增加、凝血时间缩短、大量纤维蛋白形成,从而促进肿瘤相关血栓形成[33]
肿瘤相关成纤维细胞(CAFs)是TME中活化的成纤维细胞,可分泌多种细胞因子,在肿瘤的发生发展中发挥着重要作用[34]。Takesue等[35]通过体内及体外实验证实,NETs可作用于胰腺导管腺癌中的CAFs,从而促进肝转移,而NETs抑制剂可减少肿瘤组织中CAFs的募集,进而抑制肝转移。Munir等[20]进一步研究发现,在黑色素瘤及胰腺癌中,这种作用是CAFs产生的淀粉样蛋白β介导的,抑制NETs或淀粉样蛋白β均能抑制肿瘤的生长,并恢复中性粒细胞的抗肿瘤活性。
NETs还可通过调控TME中的细胞外基质及细胞因子而影响肿瘤的进展。一方面,Albrengues等[11]发现,持续的肺部炎症会导致在休眠癌细胞周围形成NETs,作为NETs的重要组成部分,NE及MMP9依次作用于细胞外基质中的层黏连蛋白-111,使其暴露出一个新的表位,该表位能与肿瘤细胞表面的整合素α3β1结合,从而使休眠的肿瘤细胞苏醒,进一步促进肿瘤的复发与转移。另一方面,在胰腺癌中,胰腺癌细胞可产生IL-17,后者通过募集中性粒细胞诱导NETs的产生;抑制IL-17可增强免疫检查点抑制剂(如PD-1、CTLA4)的敏感性,而抑制中性粒细胞或PAD4依赖的NETosis能产生类似的效应[36]
NETs可作为多种肿瘤的标志物,与肿瘤的分期、分型密切相关。Rayes等[24]研究发现,NETs水平在肺癌、胃癌及食管癌患者中明显升高,晚期患者明显高于早期患者,且仅有肿瘤也可诱导NETosis,不依赖于手术应激及炎症反应。Zhang等[15]发现,外周血NETs水平在以下人群中从高到低依次为:胃腺癌晚期患者、胃腺癌术前患者、胃腺癌术后患者、胃良性病变患者、健康志愿者;且NETs水平与肿瘤的分型及预后相关。Park等[10]在人乳腺癌组织标本中检测到NETs,其中三阴性乳腺癌中表达量最高,Luminal型乳腺癌中表达量很少,甚至不表达。还有研究发现,NETs广泛存在于乳腺癌及结肠癌患者的肝转移病灶中,而跨膜蛋白CCDC25作为存在于肿瘤细胞表面的NETs DNA受体,可增强肿瘤细胞的活性,促进肿瘤的转移[26]。血清中NETs的水平及肿瘤细胞表面CCDC25的表达量可作为生物标志物,特异性地预测早期乳腺癌患者肝转移的风险。因此,NETs作为生物标志物在肿瘤的分期、分型及预测肿瘤进展和预后方面将发挥重要的作用。
虽然目前尚无临床试验评估靶向NETs对于肿瘤发生、发展、转移及复发的影响,但已有研究提示抑制NETs对肿瘤的早期防治具有重要意义。van der Windt等[37]发现,抑制NETs可使非酒精性脂肪肝诱发的肝癌发生率降低50%。同样地,使用NE抑制剂可以明显抑制NETs形成,阻止肿瘤细胞与肝毛细血管窦及肺毛细血管网内皮细胞的黏附,进而减少肿瘤细胞向肝脏与肺部转移[27]。另外,DNase Ⅰ或PAD4抑制剂可抑制NETs的形成,进一步抑制肿瘤的发生发展[9-13,28],且PAD4抑制剂或DNase Ⅰ也可减轻NETs介导的肿瘤相关肾功能损伤[38]。目前,NE抑制剂及DNase Ⅰ已用于其他呼吸系统疾病的治疗[39]。以上研究提示,NETs作为肿瘤治疗的靶点可能会有很好的应用前景,当然,考虑到不同肿瘤的特异性、不同治疗方式的影响及肿瘤患者与动物模型的差异,将NETs抑制剂真正应用于临床还须谨慎。
中性粒细胞一直以来被认为是人体重要的防御机制,可通过NETs诱捕并杀灭有害病原体。目前研究发现,NETs在多种肿瘤中高表达,TME中的成分能够刺激NETs的形成,而NETs也能通过影响TME中的肿瘤细胞、免疫细胞、内皮细胞、成纤维细胞及细胞外基质等促进肿瘤增殖、侵袭及转移。以上研究结果提示,NETs作为预测肿瘤进展及预后的标志物,以及潜在治疗靶点将有很好的应用前景。但由于受到肿瘤类型、治疗方式及肿瘤患者与动物模型差异的影响,具体的作用机制还需进一步探讨。
  • 国家自然科学基金(81770525)
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2022年第47卷第4期
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doi: 10.11855/j.issn.0577-7402.2022.04.0407
  • 接收时间:2021-03-31
  • 首发时间:2025-12-17
  • 出版时间:2022-04-28
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  • 收稿日期:2021-03-31
  • 录用日期:2021-06-15
基金
National Natural Science Foundation of China(81770525)
国家自然科学基金(81770525)
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    兰州大学第二医院消化内科,兰州 730030

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张德奎,E-mail:
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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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