circZFAND6调控三阴型乳腺癌TAMs向M2型极化的机制  

Mechanism of circZFAND6 in regulating the polarization of TAMs to M2 type in triple negative breast cancer

作　　者：刘睿涵 叶熹罡[2] 王守亮 张孟利 毛思怡 余建武 李舜[1,2] 谭维格[1,2] LIU Ruihan;YE Xigang;WANG Shouliang;ZHANG Mengli;MAO Siyi;YU Jianwu;LI Shun;TAN Weige(Department of Breast Surgery,the First Affiliated Hospital of Guangzhou Medical University,Guangzhou 510080,China;Guangzhou Medical University,Guangzhou 510080,China)

机构地区：[1]广州医科大学附属第一医院乳腺外科,广州510080 [2]广州医科大学,广州510080

出　　处：《华夏医学》2024年第5期1-7,共7页Acta Medicinae Sinica

基　　金：国家自然科学基金委青年科学基金项目(81902948);广州市科技基础与应用基础研究项目(202201010029)。

摘　　要：目的探讨circZFAND6调控三阴型乳腺癌(TNBC)肿瘤相关巨噬细胞(TAMs)向M2型极化的调控机制。方法构建用Luciferase重组病毒感染和标记细胞系;用CRISPR/Cas9自带EGFP系统靶向敲低circZFAND6的表达;细胞生物学功能实验等检测circZFAND6对TNBC细胞增殖、克隆形成及侵袭能力的影响。qRT-PCR、免疫组化、Western blot检测肿瘤组织或细胞中免疫相关基因和蛋白的表达情况。结果circZFAND6在TNBC肿瘤组织中表达上调,可促进乳腺癌细胞增殖、侵袭等恶性转化。而circZFAND6高表达的肿瘤组织中CD3+T细胞、CD8+T细胞浸润明显减少,而CD163标记的M2型TAMs明显增多。过表达circZFAND6可上调c-MYC和CD47蛋白水平,从而促进免疫T细胞的耗竭,并推动TAMs向M2型极化。结论在TNBC中,过表达circZFAND6可促进肿瘤免疫微环境中TAMs向M2型极化以及T细胞的耗竭。Objective To explore the mechanism of circZFAND6 in regulating the polarization of tumorassociated macrophages(TAMs)to M2 type in triple-negative breast cancer(TNBC).Methods Luciferase recombinant virus was used to infect and label cell lines.circZFAND6 expressed was knocked out using CRISPR/Cas9 self-contained EGFP system.The effects of circZFAND6 on the proliferation,clonogenesis and invasion of TNBC cells were determined by cell counting,clonogenesis and Transwell invasion assay.The expression of immune-related genes and proteins in tumor tissues or cells were detected by qRT-PCR,immunohistochemistry and Western blot.Results circZFAND6 is up-regulated in TNBC tumor tissues,which can promote malignant transformation such as proliferation and invasion of breast cancer cells.In addition,the expression of circZFAND6 increased in immunotherapy-resistant TNBC tumor tissues,while the infiltration of CD3+T cells and CD8+T cells in tumor tissues of patients with high expression of circZFAND6 decreased significantly,while the number of CD163-labeled M2 TAMs increased significantly.At the same time,overexpression of circZFAND6 may up-regulate c-MYC and CD47 protein levels in breast cancer cells,thus promoting the depletion of immune T cells and promoting the polarization of TAMs to M2 type.Conclusion In TNBC,circZFAND6 is closely related to the immunotherapy and immune microenvironment of TNBC.The high expression of circZFAND6 can promote the polarization of TAMs to M2 type and the depletion of T cells in the tumor immune microenvironment.

关 键 词：三阴型乳腺癌 肿瘤相关巨噬细胞 circZFAND6 M2型极化 肿瘤免疫微环境 

分 类 号：R737.9[医药卫生—肿瘤]