CCAAT/增强子结合蛋白δ通过调控巨噬细胞向M1型极化抑制肝癌侵袭转移  

CCAAT/enhancer binding proteinδinhibits invasion and metastasis of liver cancer by regulating MI type macrophages polarization

作　　者：李梦 熊永福 黄徐建 陈泰安 李敬东[1] Li Meng;Xiong Yongfu;Huang Xujian;Chen Taian;Li Jingdong(Department of Hepatology,Affiliated Hospital of North Sichuan Medical College,Hepatobiliary and Intestine Research Institute,North Sichuan Medical Collge,Nanchong 637000,China)

机构地区：[1]川北医学院附属医院肝胆外科,川北医学院肝胆胰肠疾病研究所,南充637000

出　　处：《中华肝脏病杂志》2021年第8期794-798,共5页Chinese Journal of Hepatology

基　　金：四川省科技计划项目(2019YJ0384)。

摘　　要：目的探讨CCAAT/增强子结合蛋白δ(CEBPD)对巨噬细胞极化的调控及通过巨噬细胞对肝癌细胞侵袭转移、凋亡的影响。方法用慢病毒转染技术构建敲减CEBPD(shCEBPD)及阴性对照shNC的THP-1稳定转染细胞。用佛波醇12-十四酸酯13-乙酸酯(PMA)将转染后的THP-1细胞诱导为巨噬细胞,脂多糖(LPS)和干扰素Y(IFNy)进一步将巨噬细胞向MI型极化诱导。实时荧光定量聚合酶链反应(qRT-PCR)检测MI型巨噬细胞相关基因白细胞介素1β(IL-1β)、IL-6、肿瘤坏死因子α(TNFα)及诱导型一氧化氮合酶(iNOS)的mRNA表达水平,流式细胞术检测MI型巨噬细胞特异表面标志物CD80表达水平。用Transwell非接触式共培养小皿将经MI型诱导后的巨噬细胞与肝癌MHCC97H细胞进行共培养。在共培养条件下,通过Transwell和划痕实验检测MHCC97H的侵袭转移能力,流式细胞术检测MHCC97H细胞的凋亡情况。组间数据比较采用t检验分析。结果敲减CEBPD后,THP-1来源巨噬细胞中MI型巨噬细胞标志基因iNOS、TNFα、IL-6及IL-1β的mRNA表达水平降低,MI型巨噬细胞表面特异标志物CD80表达减少(23.7%±2.1%与62.5%±2.0%,t=9.58,P<0.05)。将shCEBPD组和shNC组的THP-1分别与MHCC97H进行共培养,与shNC组相比,shCEBPD组共培养的MHCC97H细胞侵袭能力[(158.0±3.5)个与(75.0±4.5)个,t=39.87,P<0.01]和转移能力(54.6%±1.5%与24.3%±1.0%,t=61.42,P<0.01)增强、凋亡率降低[(9.4%±1.0%)与(23.7%±1.2%),t=12.68,P<0.01]。结论CEBPD通过促进巨噬细胞M1型极化抑制肝癌侵袭转移,并增加肝癌细胞凋亡。Objective To explore the regulation of macrophage polarization and its effects on liver cancer invasion,metastasis and apoptosis by CCAAT/enhancer binding proteinδ(CEBPD).Methods THP-1 stable transfected cells with knockdown CEBPD(shCEBPD)and negative control shNC were constructed by lentviral transfection technique.THP-I transfected cells were induced into macrophages,lipopolysaccharide(LPS)and interferonγ(IFNγ)by phorbol l2-tetradecanoate 13-acetate(PMA),and then the polarized macrophages were further induced to MI type.The quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect MI type macrophage related interleukin 1β(IL-1β)genes,IL-6,tumor necrosis factor α(TNFα),and inducible nitric oxide synthase(iNOS)mRNA expression level.Flow cytometry was used to detect MI macrophage-specific surface marker CD80 expression levels.Ml-induced macrophages were co-cultured with liver cancer MHCC97H cells using Transwell non-contact small sized co-culture dishes.MHCC97H cells invasion and metastasis were detected by Transwell and scratch assay under co-culture conditions,and the MHCC97H cells apoptosis was detected by flow cytometry.Results The mRNA expression levels of MI macrophage marker genes iNOS,TNFα,IL-6 and IL-1β in THP-1 derived macrophages were decreased after CEBPD knockdown.MI macrophage-specific surface marker CD80 expression levels were decreased(23.7%±2.1%and 62.5%±2.0%,t=9.58,P<0.05).THP-1 were co-cultured with MHCC97H in shCEBPD and shNC group,respectively.Compared with shNC group,the invasion[(158.0±3.5)and(75.0±4.5),t=39.87,P<0.01]and metastatic ability(54.6%±1.5%and 24.3%±1.0%,P<0.01)of MHCC97H cells co-cultured in shCEBPD group were stronger and the apoptosis rate was reduced[(9.4%±1.0%)vs.(23.7%±1.2%),t=12.68,P<0.01].Conclusion CEBPD can inhibit the invasion and metastasis and increase the apoptosis by amplifying MI type macrophages polarization in liver cancer cells.

关 键 词：肝细胞癌 巨噬细胞 M1极化 CCAAT/增强子结合蛋白δ 

分 类 号：R735.7[医药卫生—肿瘤]