吉非替尼耐药肺癌细胞(HCC827/GR)分泌CXCL8影响肿瘤微环境中Treg细胞浸润  

Gefitinib-resistant lung cancer cells(HCC827/GR)to affect Treg cell infiltration in tumor microenvironment via secrete CXCL8

作　　者：蔡为为 温清国 庞艳波 卢巍 CAI Wei-wei;WEN Qing-guo;PANG Yan-bo;LU Wei(Department of Respiratory and Critical Care Medicine,Northern Theater Command General Hospital,Shenyang 110016,China;Department of Respiratory Medicine,Northern Theater Command General Hospital,Shenyang 110016,China;Department of Clinical Diagnosis,Northern Theater Command General Hospital,Shenyang 110016,China)

机构地区：[1]解放军北部战区总医院呼吸与危重症医学科,辽宁沈阳110016 [2]解放军北部战区总医院呼吸科,辽宁沈阳110016 [3]解放军北部战区总医院干诊科,辽宁沈阳110016

出　　处：《解剖科学进展》2024年第5期491-494,共4页Progress of Anatomical Sciences

基　　金：全军“十二五”科研基金项目(CSY14C005)。

摘　　要：目的探究吉非替尼耐药肺癌HCC827/GR细胞对与其共培养的T细胞Treg细胞极化的影响,并分析其机制。方法采用药物浓度递增法建立吉非替尼耐药的肺癌HCC827/GR细胞,将HCC827/GR细胞与CD4+T细胞共培养,采用流式细胞术检测CD4+T细胞中Treg细胞比例。RT-qPCR和ELISA检测HCC827/GR细胞CXCL8表达及分泌情况。将敲减CXCL8慢病毒转染HCC827/GR细胞,ELISA法检测HCC827/GR细胞CXCL8分泌情况,流式细胞术检测HCC827/GR细胞共培养的CD4+T细胞中Treg细胞比例,Western blot检测HCC827/GR细胞共培养的CD4+T细胞中CXCR1和TGF-β蛋白表达情况。结果与HCC827/GR细胞共培养的CD4+T细胞中CD4+CD25+Foxp3+细胞的比例较HCC827细胞明显升高,HCC827/GR细胞中CXCL8 mRNA表达以及细胞培养基上清液中CXCL8水平均高于HCC827细胞。敲减CXCL8降低HCC827/GR细胞培养基上清液中CXCL8水平,并降低与HCC827/GR细胞共培养的CD4+T细胞中CD4+CD25+Foxp3+细胞,下调CD4+T细胞中CXCR1和TGF-β蛋白表达。结论吉非替尼耐药的NSCLC细胞促进肿瘤微环境中的Treg细胞极化,其机制可能与CXCL8分泌增加介导的T细胞中CXCR1/TGF-β信号通路激活有关。Objective To explore the effect of gefitinib-resistant lung cancer HCC827/GR cells on the polarization of T cell Treg cells co-cultured with it,and analyze its mechanism.Methods Gefitinib-resistant lung cancer HCC827/GR cells were established by increasing drug concentration.HCC827/GR cells were co-cultured with CD4+T cells,and the proportion of Treg cells in CD4+T cells was detected by flow cytometry.RT-qPCR and ELISA were used to detect the expression and secretion of CXCL8 in HCC827/GR cells.HCC827/GR cells were transfected with CXCL8 knockdown lentivirus,and the secretion of CXCL8 in HCC827/GR cells was detected by ELISA,the proportion of Treg cells in CD4+T cells co-cultured with HCC827/GR cells was detected by flow cytometry,and the expressions of CXCR1 and TGF-βin CD4+T cells co cultured with HCC827/GR cells were detected by Western blot.Results The proportion of CD4+CD25+Foxp3+cells in CD4+T cells co-cultured with HCC827/GR cells was significantly higher than that of HCC827 cells.The expression of CXCL8 mRNA in HCC827/GR cells and the level of CXCL8 in cell culture medium supernatant were higher than those of HCC827 cells.Knockdown of CXCL8 decreased the level of CXCL8 in the supernatant of HCC827/GR cell culture medium,decreased CD4+CD25+Foxp3+cells in CD4+T cells co-cultured with HCC827/GR cells,and down-regulated the expressions of CXCR1 and TGF-βin CD4+T cells.Conclusion Gefitinib-resistant NSCLC cells promote the polarization of Treg cells in tumor microenvironment,and its mechanism may be related to the activation of CXCR1/TGF-βsignal pathway in T cells mediated by the increase of CXCL8 secretion.

关 键 词：肺癌 吉非替尼耐药 肿瘤微环境 Treg细胞 CXCL8 CXCR1/TGF-β信号通路 

分 类 号：R734.2[医药卫生—肿瘤]