PAD4介导中性粒细胞胞外诱捕网促甲状腺癌细胞增殖、侵袭的机制研究  

Mechanism of PAD4-mediated neutrophil extracellular traps promoting proliferation and invasion of thyroid cancer cells

作　　者：雪克来提·库尔班[1] 苏鹏程[1] 阿米娜·阿不都热合曼 Xuekelaiti·Kuerban;SU Peng-cheng;Amina·Abudureheman(Department of Breast and Thyroid,Xinjiang Uygur Autonomous Region People's Hospital,Urumqi Xinjiang 830001,China;Health Management Center,Xinjiang Uygur Autonomous Region People's Hospital,Urumqi Xinjiang 830001,China)

机构地区：[1]新疆维吾尔自治区人民医院乳腺甲状腺科,新疆乌鲁木齐830001 [2]新疆维吾尔自治区人民医院健康管理中心,新疆乌鲁木齐830001

出　　处：《局解手术学杂志》2024年第8期701-706,共6页Journal of Regional Anatomy and Operative Surgery

基　　金：国家重点研发计划研究项目(2018YFC1708003)。

摘　　要：目的探究肽酰基精氨酸脱亚胺酶4(PAD4)介导的中性粒细胞胞外诱捕网(NETs)在甲状腺癌细胞增殖、侵袭和细胞周期中的作用机制以及临床相关性研究。方法收集60例甲状腺癌患者和60例健康受试者的外周血,ELISA检测血清中PAD4和组蛋白H3瓜氨酸化(H3cit)水平,并分析甲状腺癌患者血清中PAD4和H3cit水平与疾病进展的相关性。提取甲状腺癌患者和健康受试者外周血中性粒细胞,qRT-PCR检测PAD4 mRNA表达水平,离子霉素刺激和绿菁染色检测NETs形成水平。将NETs、GSK484与人甲状腺癌细胞株CAL-62共孵育,分为Control组、NETs组和NETs+GSK484组。采用CCK-8法、Transwell法和流式细胞术分别检测细胞增殖和侵袭水平以及细胞周期进展。结果与健康受试者相比,甲状腺癌患者血清中PAD4和H3cit水平升高(P<0.05),且PAD4和H3cit水平呈正相关(P<0.05)。甲状腺癌患者的PAD4和H3cit水平均与TNM分期有关(P<0.05)。与健康受试者相比,甲状腺癌患者外周血中性粒细胞PAD4 mRNA表达及NETs形成水平升高(P<0.05)。与Control组相比,NETs组细胞增殖、侵袭水平升高(P<0.05),S期细胞比例升高(P<0.05),G1期细胞比例降低(P<0.05)。GSK484可抑制NETs形成。与NETs组相比,NETs+GSK484组细胞增殖、侵袭水平降低(P<0.05),S期细胞比例降低(P<0.05),G1期细胞比例升高(P<0.05)。结论PAD4介导的NETs通过激活细胞周期进展,促进甲状腺癌细胞增殖和侵袭,加速甲状腺癌疾病进展。Objective To investigate the mechanism and clinical relevance of peptidyl arginine deiminase 4(PAD4)-mediated neutrophil extracellular traps(NETs)in the proliferation,invasion,and cell cycle of thyroid cancer cells.Methods Peripheral blood of 60 thyroid cancer patients and 60 healthy subjects were collected.Serum PAD4 and citrullinated histone H3(H3cit)levels were detected by ELISA,and the correlations between levels of serum PAD4 and H3cit in thyroid cancer patients and disease progression were analyzed.Neutrophils were extracted from peripheral blood of thyroid cancer patients and healthy subjects,PAD4 mRNA expression levels were detected by qRT-PCR,NETs formation level was detected by ionomycin stimulation and green cyanine staining.Human thyroid cancer cell line CAL-62 cells were co-cultured with NETs and GSK484,and divided into the Control group,the NETs group and the NETs+GSK484 group.CCK-8 assay,Transwell assay,and flow cytometry were used to evaluate the levels of cell proliferation and invasion,and the cell cycle progression,respectively.Results Compared with healthy subjects,the levels of serum PAD4 and H3cit in thyroid cancer patients were increased(P<0.05),and the levels of PAD4 and H3cit were positively correlated(P<0.05).The levels of PAD4 and H3cit in thyroid cancer patients were correlated with TNM stage(P<0.05).Compared with healthy subjects,the expression of PAD4 mRNA and the NETs formation level of peripheral blood neutrophils in thyroid cancer patients increased(P<0.05).Compared with the Control group,the levels of cell proliferation and invasion,and the proportion of S-phase cells increased(P<0.05),while the proportion of G1-phase cells decreased in the NETs group(P<0.05).GSK484 can inhibit the formation of NETs.Compared with the NETs group,the levels of cell proliferation and invasion,and the proportion of S-phase cells decreased(P<0.05),while the proportion of G1-phase cells increased in the NETs+GSK484 group(P<0.05).Conclusion PAD4-mediated NETs promote the proliferation and invasion of th

关 键 词：甲状腺癌 肽酰基精氨酸脱亚胺酶4 中性粒细胞胞外诱捕网 组蛋白H3瓜氨酸化 

分 类 号：R736.1[医药卫生—肿瘤]