G-CSF刺激荷瘤小鼠骨髓中性粒细胞产生中性粒细胞胞外陷阱促进U14小鼠宫颈癌细胞成瘤作用的研究

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英文篇名：G-CSF Produces Neutrophil Extracellular Traps in Murine Tumor-bearing Bone Marrow Neutrophils and Induces the Tumor-promoting Effect of U14 Murine Cervical Cancer Cell Line 作者：颜彬 ; 李艳丽 ; 马全富 ; 周志刚 ; 郭玉琳 ; 戴璇 ; 高晗 ; 吴绪峰 英文作者：Yan Bin;Li Yanli;Ma Quanfu;Department of Gynecologic Oncology,Hubei Maternity and Child Health Hospital; 关键词：宫颈癌 ; 粒细胞集落刺激因子 ; 小鼠骨髓中性粒细胞 ; 中性粒细胞胞外陷阱 ; 促瘤作用 英文关键词：cervical cancer;;granulocyte colony-stimulating factor;;mouse bone marrow neutrophils;;neutrophil extracellular traps;;tumor growth 中文刊名：TJYX 英文刊名：Acta Medicinae Universitatis Scientiae et Technologiae Huazhong 机构：湖北省妇幼保健院肿瘤妇科; 出版日期：2019-08-06 出版单位：华中科技大学学报(医学版) 年：2019 期：v.48 基金：国家自然科学基金青年基金项目(No.81502265);; 湖北省卫健委科研项目(No.WJ2019M228);; 湖北省科技厅创新团队专项(No.2017CKC891) 语种：中文; 页：TJYX201904003 页数：5 CN：04 ISSN：42-1678/R 分类号：18-22

摘要

目的研究粒细胞集落刺激因子(G-CSF)作用下荷瘤小鼠骨髓中性粒细胞(TBM-PMN)产生中性粒细胞胞外陷阱(NETs)对U14宫颈癌细胞成瘤、血管生成及凋亡的作用。方法 Percoll密度梯度离心法分离获取TBM-PMN,在体外实验中用G-CSF刺激TBM-PMN产生NETs,用流式细胞术检测NETs的生成。体内实验研究U14肿瘤细胞与TBM-PMN混合成瘤后,用G-CSFR干扰G-CSF作用或DNA酶Ⅰ抑制NETs形成,研究其对U14肿瘤细胞成瘤的影响,免疫组织化学法检测其对肿瘤组织中血管生成的影响。在体外实验中将U14肿瘤细胞与TBM-PMN共培养,用anti-G-CSF阻断G-CSF作用或DNA酶Ⅰ抑制NETs形成,流式细胞术检测其对U14肿瘤细胞凋亡的影响。结果 G-CSF刺激TBM-PMN产生NETs。在体内实验中TBM-PMN有促进肿瘤生长的作用,加入G-CSFR或DNA酶Ⅰ后其促肿瘤生长作用及促血管生成作用减弱。在体外TBM-PMN与U14肿瘤细胞共培养体系中,用anti-G-CSF阻断G-CSF作用或者用DNA酶Ⅰ抑制NETs形成后其抑制凋亡的能力可部分恢复。结论 G-CSF作用下小鼠TBM-PMN能够产生NETs,其对U14小鼠宫颈癌细胞株具有促瘤作用。
        Objective To investigate the effects of neutrophil extracellular traps(NETs)produced by granulocyte colony-stimulating factor(G-CSF)stimulating mouse tumor-bearing bone marrow neutrophils(TBM-PMN)on the tumor growth,angiogenesis and apoptosis of U14 murine cervical cancer cell line.Methods TBM-PMN was isolated by Percoll density gradient centrifugation.In in vitro experiment,G-CSF stimulated TBM-PMN to generate NETs,and flow cytometer was used to detect the formation of NETs.In in vivo experiments,U14 tumor cells were co-inoculated with TBM-PMN to form tumors,using G-CSFR to block the effect of G-CSF or DNase Ⅰ to inhibit the NETs formation,and their effects on U14 tumor cell formation were investigated,and their effects on angiogenesis in tumor tissues were detected by immunohistochemical method.In in vitro experiments,U14 tumor cells were co-cultured with TBM-PMN,with anti-G-CSF blocking the effect of G-CSF or DNase Ⅰ inhibiting NETs formation,and their effects on U14 tumor cell apoptosis were detected by flow cytometry.Results G-CSF stimulated TBM-PMN to produce NETs.In vivo TBM-PMN promoted tumor growth and angiogenesis of U14 cell line and its promoting effects were attenuated after adding G-CSFR to block G-CSF or DNase Ⅰ to inhibit NETs formation.In vitro co-culture system of TBM-PMN and U14 cervical cancer line,with the use of anti-G-CSF antibody to block G-CSF or DNase Ⅰ to inhibit NETs formation,the ability of TBM-PMN inhibiting apoptosis of U14 cell line could be partly recovered.Conclusion Mouse TBM-PMN can produce NETs under G-CSF,which has a tumor-promoting effect on U14 mouse cervical cancer cell line.

引文

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