PAD4介导H3cit促结肠癌细胞增殖、侵袭和上皮间质转化的机制研究  

Mechanism research of PAD4-mediated H3cit to promotes the proliferation,invasion and epidermal-mesenchymal transition of colon cancer cells

作　　者：王永琦[1] 刘丽杰 何冬雷[1] 陈勇[1] WANG Yong-qi;LIU Li-jie;HE Dong-lei;CHEN Yong(Department of Gastrointestinal Oncology,the First Affiliated Hospital of Hainan Medical College,Haikou Hainan 570102,China)

机构地区：[1]海南医学院第一附属医院胃肠肿瘤外科,海南海口570102

出　　处：《局解手术学杂志》2024年第4期281-287,共7页Journal of Regional Anatomy and Operative Surgery

基　　金：海南省卫生健康行业科研项目(21A200052)。

摘　　要：目的 探究肽基精氨酸脱亚胺酶4(PAD4)介导的瓜氨酸化组蛋白H3(H3cit)在结肠癌细胞增殖、侵袭和上皮间质转化中的作用及其相关机制。方法 收集我院30例结肠癌患者的血清和30例健康受试者的血清,ELISA检测血清中PAD4和H3cit水平,并分析两者相关性。将oe-NC、过表达PAD4(oe-PAD4)和sh-NC、sh-NLRP3载体转染至人结肠癌SW480细胞中,并设为oe-NC组、oe-PAD4组、oe-PAD4+sh-NC组、oe-PAD4+sh-NLRP3组。采用qRT-PCR检测PAD4、NLRP3、IL-1β、IL-18基因表达水平;Western blot检测PAD4、H3cit、NLRP3、E-cadherin、N-cadherin蛋白表达;CCK-8检测细胞增殖水平;Transwell检测细胞侵袭水平;ChIP-qPCR检测H3cit在NLRP3转录起始位点(TSS)的富集水平。结果 与健康受试者比较,结肠癌患者血清中PAD4和H3cit水平均升高,且两者呈正相关(P<0.05)。与oe-NC组比较,oe-PAD4组PAD4、NLRP3、IL-1β、IL-18基因表达水平升高,PAD4、H3cit、NLRP3、N-cadherin蛋白表达升高,E-cadherin蛋白表达降低,细胞增殖、侵袭水平升高,H3cit在NLRP3 TSS区富集增加(P<0.05);与oe-PAD4+sh-NC组比较,oe-PAD4+sh-NLRP3组NLRP3、IL-1β、IL-18基因表达水平降低,NLRP3和N-cadherin蛋白表达降低,E-cadherin蛋白表达升高,细胞增殖、侵袭水平降低(P<0.05)。结论 结肠癌患者体内PAD4和H3cit水平升高,PAD4介导的H3cit通过转录调控NLRP3的表达,促进结肠癌细胞增殖、侵袭和上皮间质转化。Objective To explore the role and mechanism of peptidyl arginine deiminase 4(PAD4)-mediated citrullinated histones H3(H3cit)in the proliferation,invasion and epidermal-mesenchymal transition of colon cancer cells.Methods Serum samples were collected from 30 colon cancer patients and 30 healthy subjects in our hospital.The levels of PAD4 and H3cit in serum were detected by ELISA,and the correlation between the two was analyzed.oe-NC,overexpressed PAD4(oe-PAD4),sh-NC and sh-NLRP3 vectors were transfected into human colon cancer SW480 cells and divided into the oe-NC group,the oe-PAD4 group,the oe-PAD4+sh-NC group,and the oe-PAD4+sh-NLRP3 group.The gene expression levels of PAD4,NLRP3,IL-1βand IL-18 were detected by qRT-PCR,the protein expression of PAD4,H3cit,NLRP3,E-cadherin and N-cadherin were detected by Western blot.The cell proliferation level was detected by CCK-8,and the level of cell invasion was detected by Transwell assay.The level of H3cit enrichment in NLRP3 transcription start sites(TSS)was detected by ChIP-qPCR.Results Compared with healthy subjects,the levels of PAD4 and H3cit in serum of colon cancer patients were increased,and there was a positive correlation between the two(P<0.05).Compared with the oe-NC group,the gene expression levels of PAD4,NLRP3,IL-1βand IL-18 were increased,the protein expression of PAD4,H3cit,NLRP3 and N-cadherin were increased,the protein expression of E-cadherin was decreased,the cell proliferation and invasion levels were increased,and the enrichment of H3cit in NLRP3 TSS zone was increased in the oe-PAD4 group(P<0.05).Compared with the oe-PAD4+sh-NC group,the gene expression levels of NLRP3,IL-1βand IL-18 were decreased,the protein expression of NLRP3 and N-cadherin were decreased,the protein expression of E-cadherin was increased,and the cell proliferation and invasion levels were decreased in the oe-PAD4+sh-NLRP3 group(P<0.05).Conclusion The levels of PAD4 and H3cit in colon patients are increased,and the PAD4-mediated H3cit promotes the proliferation,invasion and

关 键 词：结肠癌 肽基精氨酸脱亚胺酶4 瓜氨酸化组蛋白H3 上皮间质转化 转录调控 

分 类 号：R735.35[医药卫生—肿瘤]