miR-381在HCC患者肿瘤组织、外周血中表达及过表达对巨噬细胞极化模式调节作用  

Expression of miR-381 in tumor tissues and peripheral blood of HCC patients and the regulatory effect of its overexpression on polarization pattern of macrophages

作　　者：吴书志 王菁 WU Shuzhi;WANG Jing(Shandong Center for Disease Control and Prevention,Jinan 250013,China;不详)

机构地区：[1]山东省疾病预防控制中心,济南250013 [2]山东第一医科大学附属中心医院医学实验诊断中心

出　　处：《山东医药》2023年第33期1-6,共6页Shandong Medical Journal

基　　金：济南市科技计划项目(202019046);济南市卫生健康委员会科技计划项目(2020-4-20)。

摘　　要：目的观察miR-381在肝细胞癌(HCC)患者肿瘤组织、外周血和体外培养巨噬细胞中的表达变化,并探讨miR-381过表达对巨噬细胞极化模式的调节作用。方法HCC患者40例,术中留取肿瘤组织与癌旁组织,采用RT-PCR法检测miR-381、一氧化氮合酶(iNOS,M1型巨噬细胞标记物)mRNA、精氨酸酶1(Arg-1,M2型巨噬细胞标记物)mRNA,采用Western Blotting法检测iNOS、Arg-1蛋白。抽取HCC患者和健康志愿者外周静脉血,采用RT-PCR法检测miR-381、CD86(M1型巨噬细胞标记物)mRNA、CD163(M2型巨噬细胞标记物)mRNA,采用流式细胞法检测CD86、CD163蛋白。取人髓系白血病单核细胞THP-1,分别诱导分化成M0型巨噬细胞(M0组)、M2型巨噬细胞(M2组)、肿瘤相关巨噬细胞(TAMs,CM组),采用RT-PCR法检测各组细胞miR-381、iNOS mRNA、Arg-1 mRNA、CD86 mRNA、CD163 mRNA,采用Western Blotting法和流式细胞法检测iNOS、Arg-1、CD86、CD163蛋白。取M0型巨噬细胞,分别转染miR-381模拟物(miR-381 mimic)、对照模拟物(mimic NC),记为miR-381 mimic组、mimic NC组,建立miR-381过表达细胞模型,采用RT-PCR法检测各组细胞miR-381、iNOS mRNA、Arg-1 mRNA、CD86 mRNA、CD163 mRNA,采用Western Blotting法和流式细胞法检测iNOS、Arg-1、CD86、CD163蛋白。结果HCC患者肿瘤组织中miR-381、iNOS mRNA和蛋白相对表达量低于癌旁组织,Arg-1 mRNA和蛋白相对表达量高于癌旁组织(P均<0.05)。HCC患者外周血巨噬细胞中miR-381、CD86 mRNA和蛋白表达水平低于健康志愿者,CD163 mRNA和蛋白表达水平高于健康志愿者(P均<0.05)。CM组巨噬细胞中miR-381、iNOS mRNA、Arg-1 mRNA、CD86 mRNA、CD163 mRNA和蛋白相对表达量介于M0组、M2组之间(P均<0.05)。miR-381 mimic组巨噬细胞中miR-381、iN-OS mRNA、CD86 mRNA和蛋白表达水平均升高,而Arg-1 mRNA、CD163 mRNA和蛋白表达水平均降低(P均<0.05)。结论miR-381在HCC患者肿瘤组织、外周血和体外培养巨噬细胞中均呈低表达,过表达miR-381�Objective To observe the expression changes of miR-381 in the tumor tissue,peripheral blood,and cul⁃tured macrophages of hepatocellular carcinoma(HCC)patients,and to explore the regulatory effect of miR-381 overexpres⁃sion on polarization pattern of macrophages.Methods In 40 HCC patients,tumor tissues and adjacent tissues were col⁃lected during operation,and the mRNA levels of miR-381,nitric oxide synthase(iNOS,M1 macrophage marker)and argi⁃nase 1(Arg-1,M2 macrophage marker)were detected by RT-PCR.The iNOS and Arg-1 proteins were detected by West⁃ern blotting.Peripheral venous blood of HCC patients and healthy volunteers was collected,and mRNA of miR-381,CD86(M1 macrophage marker)and CD163(M2 macrophage marker)were determined by RT-PCR,and proteins of CD86 and CD163 were determined by flow cytometry.Human myeloid leukemia monocyte THP-1 was induced to differentiate in⁃to M0 type macrophages(M0 group),M2 type macrophages(M2 group)and tumor-associated macrophages(TAMs,CM group),respectively.In each group,miR-381,iNOS mRNA,Arg-1 mRNA,CD86 mRNA,and CD163 mRNA were mea⁃sured by RT-PCR,and iNOS,Arg-1,CD86,and CD163 proteins were measured by Western blotting and flow cytometry.M0 macrophages were transfected with miR-381 mimic and mimic NC,respectively,and were labeled as the miR-381 mimic group and mimic NC group,and we established the miR-381 overexpression cell model.RT-PCR was used to detect miR-381,iNOS mRNA,Arg-1 mRNA,CD86 mRNA,and CD163 mRNA;Western blotting and flow cytometry were used to detect iNOS,Arg-1,CD86,and CD163 proteins.Results The relative expression levels of miR-381,iN⁃OS mRNA and protein in tumor tissues of HCC patients were lower than those in the adjacent tissues,while the relative ex⁃pression levels of Arg-1 mRNA and protein were higher than those in the adjacent tissues(all P<0.05).The expression levels of miR-381 and CD86 mRNA and protein in the peripheral blood macrophages of HCC patients were lower than those of healthy volunteers,while the expression levels of CD163 mRNA a

关 键 词：肝细胞癌 miR-381 巨噬细胞 肿瘤相关巨噬细胞 M1型巨噬细胞 M2型巨噬细胞 

分 类 号：R735.7[医药卫生—肿瘤]