中性粒细胞胞外诱捕网通过上调DU145人前列腺癌细胞IL-8表达促进前列腺癌细胞增殖、 侵袭及迁移  

Neutrophil extracellular traps promote the proliferation,invasion and migration of prostate cancer cells by upregulating IL-8 expression in DU145 human prostate cancer cells

作　　者：罗后宙 陈国强 LUO Houzhou;CHEN Guoqiang(Department of Urology,Sanya Central Hospital,Sanya 572000,China)

机构地区：[1]三亚中心医院泌尿外科,海南三亚572000

出　　处：《细胞与分子免疫学杂志》2023年第3期261-267,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81660253);海南省卫生计生行业科研项目(19A300105)。

摘　　要：目的研究中性粒细胞胞外诱捕网(NET)对人前列腺癌细胞增殖、侵袭和迁移能力的影响及其相关机制。方法收集28例前列腺癌患者肿瘤组织,免疫组织化学染色法检测肿瘤及癌旁组织中白细胞介素8(IL-8)、淋巴细胞抗原6G(LY6G)、瓜氨酸化组蛋白H3(H3CIT)的表达情况。提取患者外周血中性粒细胞,体外用佛波酯(PMA)刺激形成NET。将纯化后的NET与DU145人前列腺癌细胞共孵育,通过核糖核酸测序(RNA-seq)检测NET刺激后DU145细胞上调的基因,并采用实时定量PCR和Western blot法进行验证。使用注释、可视化和整合发掘数据库(DAVID)对上调基因进行基因与基因组百科全书(KEGG)和基因本体(GO)分析。采用CCK-8法检测DU145细胞增殖能力,Transwell^(TM)实验检测细胞侵袭能力,划痕实验检测细胞迁移能力。敲低DU145细胞IL-8表达后,再次检测DU145细胞增殖、侵袭和迁移能力的改变情况。结果肿瘤组织IL-8、LY6G、H3CIT表达水平明显高于癌旁组织。NET刺激后,DU145细胞IL-8等638个基因表达上调,这些基因与磷脂酰肌醇3激酶/蛋白激酶B(PI3K/AKT)信号通路以及细胞增殖、侵袭功能相关。NET能促进DU145细胞的增殖、侵袭和迁移。沉默IL-8后,NET对DU145细胞的促增殖、侵袭和迁移能力下降。结论NET通过上调DU145细胞IL-8的表达,促进肿瘤细胞的增殖、侵袭和迁移。Objective To investigate the effects of neutrophil extracellular traps(NETs)on proliferation,migration,and invasion of human prostate cancer and its related mechanisms.Methods The expressions of interleukin-8(IL-8),lymphocyte antigen 6G(LY6G)and citrllinated histone H3(H3CIT)in 28 cases of tumor tissues and adjacent tissues were detected by immunohistochemical staining.Neutrophils were extracted from the patients and stimulated with PMA to form NETs in vitro.The up-regulated genes of DU145 cells stimulated by NETs were detected by RNA-seq,and verified by real time quantitative PCR and Western blot analysis.KEGG and GO analyses of upregulated genes were performed using the DAVID database.The proliferation,invasion and migration ability of DU145 cells was assessed by CCK-8 assay,Transwell^(TM) and scratch assay.After knockdown of IL-8 expression in DU145 cells,the changes of proliferation,invasion and migration ability of DU145 cells were tested over again.Results The expression levels of IL-8,LY6G,and H3CIT in tumor sites were significantly higher than those in adjacent tissues.After co-incubated with NETs,the expression of 638 genes including IL-8 were up-regulated in DU145 cells,which related to phosphatidylinositol 3 kinase/protein kinase B(PI3K/AKT)signaling pathway,cell proliferation and invasion.NETs can promote the proliferation,invasion and migration of DU145.After silencing IL-8,the ability of NETs to promote the proliferation,invasion and migration of DU145 was decreased.Conclusion NETs promote proliferation,migration,and invasion of DU145 by upregulating the expression of IL-8 in DU145 cells.

关 键 词：中性粒细胞胞外诱捕网(NET) 前列腺癌 DU145细胞 小干扰RNA 

分 类 号：Q279[生物学—细胞生物学] R737.25[医药卫生—肿瘤] R392-33[医药卫生—临床医学]