吉非替尼耐药肺癌HCC827/GR细胞对与其共培养的巨噬细胞M2极化的影响  

Effect and mechanism of gefitinib-resistant lung cancer HCC827/GR cells on M2 polarization of co-cultured macrophages

作　　者：温清国 蔡为为 赵睿暄 于昌萍 卢巍 WEN Qing-guo;CAI Wei-wei;ZHAO Rui-xuan;YU Chang-ping;LU Wei(Department of Respiratory and Critical Care Medicine,General Hospital of Northern Theater Command of PLA,Shenyang 110016,China)

机构地区：[1]中国人民解放军北部战区总医院呼吸与危重症医学科,辽宁沈阳110016

出　　处：《解剖科学进展》2024年第6期645-649,共5页Progress of Anatomical Sciences

基　　金：全军“十二五”科研基金(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巨噬细胞M2极化的影响及机制。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,并通过转染慢病毒建立稳定敲减IL-32的HCC827/GR细胞;将肺癌细胞与巨噬细胞共培养,采用流式细胞术检测巨噬细胞中CD11b^(+)CD206^(+)细胞比例;RT-qPCR检测巨噬细胞中IL-10和TGF-β mRNA表达以及肺癌细胞中IL-32 mRNA表达;ELISA检测巨噬细胞培养基上清中IL-10和TGF-β水平以及肺癌细胞培养基上清中IL-32水平;Western blot检测巨噬细胞中p-JAK2、JAK2、p-STAT3和STAT3蛋白表达水平。结果与HCC827/GR细胞共培养的巨噬细胞中CD11b^(+)CD206^(+)细胞比例明显升高,且巨噬细胞中IL-10和TGF-β mRNA表达及细胞培养基上清中IL-10和TGF-β水平明显升高。HCC827/GR细胞中IL-32 mRNA表达及细胞培养基上清液中IL-32水平均高于HCC827细胞。敲减IL-32降低HCC827/GR细胞培养基上清液中IL-32水平,并降低与HCC827/GR细胞共培养的巨噬细胞中CD11b^(+)CD206^(+)细胞比例,降低巨噬细胞中IL-10和TGF-β mRNA表达以及p-JAK2和p-STAT3蛋白表达,降低巨噬细胞培养基上清液中IL-10和TGF-β水平。结论吉非替尼耐药的NSCLC细胞促进TEM中的巨噬细胞M2极化,其机制与分泌IL-32介导巨噬细胞JAK2/STAT3信号通路激活有关。Objective To explore the effect and mechanism of gefitinib-resistant lung cancer HCC827/GR cells on M2 polarization of co-cultured macrophages.Methods Gefitinib-resistant lung cancer HCC827/GR cells were established by increasing drug concentration,and IL-32-knocked down HCC827/GR cells were established by transfection with lentivirus.Lung cancer cells were co-cultured with macrophages,and the proportion of CD11b^(+)CD206^(+)cells in macrophages was detected by flow cytometry.The levels of IL-10 and TGF-β mRNA in macrophages and IL-32 mRNA in lung cancer cells were detected by RT-qPCR.The levels of IL-10 and TGF-β in the supernatant of macrophage culture medium and IL-32 in the supernatant of lung cancer cell culture medium were detected by ELISA.The expressions of p-JAK2,JAK2,p-STAT3 and STAT3 in macrophages were detected by Western blot.Results The proportion of CD11b^(+)CD206^(+)cells in macrophages co-cultured with HCC827/GR cells were increased significantly,and the levels of IL-10 and TGF-β mRNA in macrophages and the levels of IL-10 and TGF-β in cell culture medium supernatant were increased significantly.The expression of IL-32 mRNA in HCC827/GR cells and the level of IL-32 in cell culture medium supernatant were higher than those in HCC827 cells.Knockdown of IL-32 decreased the level of IL-32 in the supernatant of HCC827/GR cell culture medium,decreased the proportion of CD11b^(+)CD206^(+)cells in macrophages co-cultured with HCC827/GR cells,decreased the expressions of IL-10,TGF-β mRNA and p-JAK2 and p-STAT3 proteins in macrophages,decreased the levels of IL-10 and TGF-β in the supernatant of macrophage culture medium.Conclusion Gefitinib-resistant NSCLC cells promote M2 polarization of macrophages in TEM,and its mechanism may be related to the activation of JAK2/STAT3 signaling pathway in macrophages mediated by IL-32 secretion.

关 键 词：肺癌 吉非替尼耐药 肿瘤微环境 巨噬细胞M2极化 IL-32 JAK2/STAT3信号通路 

分 类 号：R734.2[医药卫生—肿瘤]