结肠癌来源外泌体通过HMGB1诱导中性粒细胞活化促进结肠癌细胞增殖、侵袭和迁移  

Colon cancer cell-derived exosomes activate neutrophil through HMGB1 to promote the proliferation, invasion and migration of colon cancer cell

作　　者：周安付 谭琰 张惠晶[2] ZHOU Anfu;TAN Yan;ZHANG Huijing(Department of Gastroenterology,the First Affiliated Hospital of Hainan Medical College,Haikou 570102,China;Department of Endoscopy,the First Affiliated Hospital of China Medical University,Shenyang 110001,China)

机构地区：[1]海南医学院第一附属医院消化内科,海口570102 [2]中国医科大学附属第一医院内镜科,沈阳110001

出　　处：《中国癌症防治杂志》2022年第1期20-25,共6页CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT

基　　金：海南省卫生健康行业科研项目(19A200023)。

摘　　要：目的探讨结肠癌细胞来源外泌体在中性粒细胞和结肠癌细胞间的作用及其可能的作用机制。方法体外培养中性粒细胞和结肠癌细胞SW480,分别用sh-NC和sh-HMGB1转染SW480细胞后分离相应外泌体SW480-exo、sh-NCexo、sh-HMGB1-exo。各组外泌体分别处理中性粒细胞,同时设置PBS组;提取各组中性粒细胞上清液,再分别与SW480细胞共培养,依次记为SW480-exo+Neutrophils组、sh-NC-exo+Neutrophils组、sh-HMGB1-exo+Neutrophils组和PBS+Neutrophils组。采用Sytox green染色检测中性粒细胞胞外诱捕网(neutrophil extracellular traps,NETs)的形成情况,Western blot检测HMGB1蛋白的表达水平,CCK-8检测细胞活力,克隆形成实验检测细胞增殖情况,Transwell实验检测细胞迁移和侵袭情况。结果成功构建敲低HMGB1结肠癌细胞模型并分离SW480-exo、sh-NC-exo和sh-HMGB1-exo。与PBS组相比,SW480-exo组中性粒细胞Sytox green荧光单位和HMGB1蛋白表达水平明显升高(均P<0.05);与sh-NC-exo组相比,SW480-exo组中性粒细胞中HMGB1蛋白表达和Sytox green荧光单位差异无统计学意义(均P>0.05),但sh-HMGB1-exo组细胞中HMGB1蛋白表达和Sytox green荧光单位明显降低(均P<0.05)。SW480-exo+Neutrophils组SW480细胞活力、克隆形成数、迁移细胞数和侵袭细胞数均较PBS+Neutrophils组明显升高(均P<0.05),但与sh-NC-exo+Neutrophils组比较差异无统计学意义(均P>0.05);与sh-NC-exo+Neutrophils组相比,sh-HMGB1-exo+Neutrophils组SW480细胞活力、克隆形成数、迁移细胞数和侵袭细胞数明显降低(均P<0.05)。结论结肠癌细胞来源外泌体可能通过HMGB1诱导中性粒细胞NETs形成,从而促进细胞增殖、迁移和侵袭。Objective To investigate the function of colon cancer cell-derived exosomes in between neutrophils and colon cancer cells and its possible mechanism.Methods The neutrophils and colon cancer cells SW480 were cultured in vitro.The SW480 cells were transfected with sh-NC and sh-HMGB1,respectively,and the exosomes SW480-exo,sh-NC-exo,and sh-HMGB1-exo were isolated.Neutrophils were treated with exosomes in each group,and a PBS group was set at the same time.The neutrophil supernatant was extracted from each group,then co-cultured with SW480 cells,which were recorded as SW480-exo+Neutrophils group,sh-NC-exo+Neutrophils group,sh-HMGB1-exo+Neutrophils group and PBS+Neutrophils group,respectively.Sytox green staining was used to detect the formation of neutrophil extracellular traps(NETs);Western blot was used to detect the expression of HMGB1 protein;CCK-8 was used to detect the cell viability;Clone formation experiment was used to detect the cell proliferation;Transwell assay was used to detect the migration and invasion of cells.Results The HMGB1 knockdown colon cancer cell model was successfully constructed and SW480-exo,sh-NC-exo and sh-HMGB1-exo were isolated.Compared with the PBS group,the neutrophil Sytox green fluorescence unit and the expression of HMGB1 protein in the SW480-exo group were significantly increased(all P<0.05).Compared with the sh-NC-exo group,there was no significant difference in HMGB1 protein expression and Sytox green fluorescence unit in neutrophils in the SW480-exo group(all P>0.05),but HMGB1 protein expression and Sytox green fluorescence unit in the sh-HMGB1-exo group were significantly decreased(all P<0.05).Compared with the the PBS+Neutrophils group,the SW480 cells viability,the number of colonies,the number of migrating cells and the number of invasive cells in the SW480-exo+Neutrophils group were significantly increased(all P<0.05),but there was no significant difference compared with the sh-NC-exo+Neutrophils group(all P>0.05).Compared with the sh-NC-exo+Neutrophils group,the SW480 ce

关 键 词：结肠癌 外泌体 高迁移率族蛋白1 中性粒细胞 中性粒细胞胞外诱捕网 

分 类 号：R735.35[医药卫生—肿瘤]