肿瘤相关巨噬细胞及其分泌的CXCL1对肝癌细胞系Huh7生物学行为的影响  

Effects of TAMs and CXCL1 on the biological behavior of hepatocellular carcinoma cell line Huh7

作　　者：赵慧咏 魏昇 杨茗皓 郭子成 刘永凡 崔笑[1] Zhao Huiyong;Wei Sheng;Yang Minghao;Guo Zicheng;Liu Yongfan;Cui Xiao(Dept of General Surgery,The Second Affiliated Hospital of Anhui Medical University,Hefei 230001)

机构地区：[1]安徽医科大学第二附属医院普外科,合肥230601

出　　处：《安徽医科大学学报》2023年第3期398-404,共7页Acta Universitatis Medicinalis Anhui

基　　金：安徽高等学校自然科学研究项目(编号:KJ2017A825);安徽省自然科学基金项目(编号:1808085MH270、2008085QH406)。

摘　　要：目的探索巨噬细胞在肝癌炎症微环境中通过炎症趋化因子(C-X-C基序)配体1(CXCL1)调控肝癌细胞系Huh7进展。方法筛选肿瘤临床数据库TISIDB,分析CXCL1与肝癌患者预后的关系。免疫组化、免疫荧光染色验证肿瘤巨噬细胞与CXCL1的关系。通过佛波酯(PMA)诱导人单核细胞(THP-1)获得未分化的巨噬细胞(M0),建立与肝癌细胞共培养模型,实时荧光定量PCR法检测巨噬细胞表型的改变;细胞克隆形成实验、台盼蓝细胞活率检测肝癌细胞的增殖能力;肝癌细胞的迁移及侵袭能力使用Transwell实验检测;肝癌细胞Huh7中E-cadherin、N-cadherin、Vimentin的蛋白表达水平使用蛋白印迹法检测;酶联免疫吸附实验(ELISA法)检测细胞培养液中目标蛋白的表达。结果CXCL1高表达与肝癌患者不良预后有关,并且高表达CXCL1的肝癌组织具有更多的巨噬细胞富集。在巨噬细胞-肝癌细胞共培养模型中,两种细胞CXCL1的表达水平及共培养液中CXCL1浓度均增加。巨噬细胞M2型标志物CD163、CD206 mRNA表达水平升高。肝癌细胞Huh7的增殖、迁移及侵袭能力增强。结论巨噬细胞-肝癌细胞共培养增加肿瘤相关巨噬细胞CXCL1表达,促进肝癌细胞系Huh7进展。Objective To investigate the effects CXCL1 derived from TAMs on the progression of HCC by CXCL1 in tumor microenvironment(TME).Methods PMA induced human monocyte(THP-1)to obtain undifferentiated macrophages(M0)and then co-cultured with Huh 7 HCC cells.The biomarkers of macrophage phenotype and mRNA level of HCC were analysed by qRT-PCR.Colony formation assay,cell viability assay and transwell assay were performed to evaluate the biological alteration of HCC cells in TME.Epithelial-mesenchymal transition(EMT)molecular genetics were detected using western blot.The levels of CXCL1 in TME were measured by ELISA assay.Results High CXCL1 expression in HCC patients predicted poor prognosis.Abundant macrophages were found in CXCL1 high expression HCC tissues.In macrophages and HCC co-cultured model,significantly increasing CXCL1 expression was detected in both two cells and the biomarkers of M2 macrophages,CD163 and CD206 were elevated.The growth and migration of HCC cells were promoted.Conclusion Co-culture of macrophages with HCC cells induces macrophages pro-tumor phenotype and CXCL1 secretion which promotes the progression of HCC.

关 键 词：肝癌 肿瘤相关巨噬细胞 肿瘤微环境 CXCL1 上皮细胞-间充质转化 

分 类 号：R735.7[医药卫生—肿瘤]