HER2阳性乳腺癌细胞来源的可溶性PD-L1对巨噬细胞极化的调控作用探讨  

Regulatory effect of soluble PD-L1 derived from HER2 positive breast cancer cells on polarization of macrophages

作　　者：袁惠玲 吴丽华[1] 陈桂林[1] 黄珂铭[1] YUAN Huiling;WU Lihua;CHEN Guilin;HUANG Keming(Department of Mastopathy,Affiliated Dongguan People′s Hospital,Southern Medical University,Dongguan,Guangdong 523000,China)

机构地区：[1]南方医科大学附属东莞人民医院乳腺科,广东东莞523000

出　　处：《重庆医学》2021年第16期2701-2706,共6页Chongqing medicine

基　　金：国家自然科学基金项目(81802625);东莞市科技发展重点项目(2018507150011656)。

摘　　要：目的探讨人表皮生长因子受体2(HER2)阳性乳腺癌细胞对巨噬细胞极化的调控作用。方法ELISA检测HER2^(-)和HER2^(+)乳腺癌患者外周血和乳腺组织中可溶性程序性死亡配体1(sPD-L1)水平;流式细胞术检测乳腺组织中M1型(CD86^(+)CD206^(-)CD68^(+)/CD68^(+))和M2型(CD86-CD206^(+)CD68^(+)/CD68^(+))巨噬细胞比例;ELISA检测HER2-和HER2^(+)乳腺癌细胞系培养上清液sPD-L1水平;流式细胞术检测HER2-和HER2^(+)乳腺癌细胞系培养上清液处理后的外周血单个核细胞(PBMC)源巨噬细胞极化比例变化;实时荧光定量PCR(qRT-PCR)检测PBMC源巨噬细胞γ-干扰素(IFN-γ)、白细胞介素(IL)-1β、诱导型一氧化氮合酶(INOS)、IL-10、转化生长因子-β(TGF-β)和精氨酸酶1(ARG1)mRNA表达水平。结果HER2^(+)乳腺癌患者外周血浆和组织中sPD-L1表达水平明显高于HER2-患者(均P<0.001);HER2^(+)乳腺癌患者组织M1型巨噬细胞比例明显低于HER2-患者(P<0.001),M2型巨噬细胞比例明显高于HER2-患者(P<0.05);HER2-腺癌细胞系(SK-BR-3)和HER2^(+)腺癌细胞系(MDA-MB-453)乳腺癌细胞条件性培养基(BCM)中sPD-L1水平随培养时间增加而上升;相比于HER2-腺癌细胞系,HER2^(+)腺癌细胞系BCM中sPD-L1水平明显升高(P<0.05);HER2^(+)腺癌细胞系BCM处理的M1型和M2型巨噬细胞比例及M1/M2比值明显高于HER2-腺癌细胞系(P<0.05或P<0.001);HER2^(+)腺癌细胞系BCM处理组M1型巨噬细胞相关基因IFN-γ、IL-1β和iNOS mRNA水平明显降低(P<0.05),M2型巨噬细胞相关基因ARG1、TGF-β、IL-10 mRNA水平明显升高(P<0.001)。结论HER2^(+)和HER2-乳腺癌细胞通过sPD-L1调控肿瘤组织免疫微环境巨噬细胞极化和功能。Objective To explore the regulatory effect of human epidermal growth factor receptor 2(HER2)positive breast cancer cells on the polarization of macrophages.Methods ELISA was used to detect the level of soluble programmed death-ligand 1(sPD-L1)in the peripheral blood and breast tissue of the patients with HER2^(-)and HER2^(+)breast cancer;flow cytometry was used to detect the ratio of M1 type(CD86^(+)CD206^(-)CD68^(+)/CD68^(+))and M2 type(CD86-CD206^(+)CD68^(+)/CD68^(+))macrophages;ELISA was used to detect the sPD-L1 level in the culture supernatant of HER2-and HER2^(+)breast cancer cell lines;flow cytometry was used to detect the ratio of PBMC-derived macrophages after treated with the culture supernatant of HER2-and HER2^(+)breast cancer cell lines;quantitative real-time PCR(qRT-PCR)was used to detect theγ-interferon(IFN-γ),interleukin-1β(IL-1β),inducible nitric oxide synthase(INOS),IL-10,transforming growth factor-β(TGF-β)and arginase 1(ARG1)mRNA expression levels of PBMC-derived macrophages.Results The expression levels of sPD-L1 in plasma and tissue homogenate of HER2^(+)patients with breast cancer were significantly higher than those of HER2-patients(P<0.001);the M1 type macrophages ratio in tissue homogenate of HER2^(+)patients with breast cancer was significantly lower than that of HER2-patients,theM2 type macrophages ratio was significantly higher than that of HER2-patients(P<0.05);the sPD-L1 level was increased in breast cell conditional medium(BCM)of HER2^(+)adenocarcinoma cell line(MDA-MB-453)and HER2-adenocarcinoma cell line(SK-BR-3)in time-dependent manner;compared with HER2-adenocarcinoma cell lines,the sPD-L1 level was increased significantly in BCM of HER2^(+)adenocarcinoma cell lines(P<0.05).The proportions of M1,M2 macrophages and M1/M2 ratio treated with BCM of HER2^(+)adenocarcinoma cell lines were significantly higher than those of HER2-adenocarcinoma cell lines(P<0.05 or P<0.001);M1 type macrophages-related genes IFN-γ,IL-1βand iNOS mRNA levels were significantly decreased in HER2^(+

关 键 词：乳腺癌 人表皮生长因子受体2 可溶性程序性死亡配体1 巨噬细胞极化 

分 类 号：R737.9[医药卫生—肿瘤]