中国肺癌杂志2024，Vol.27Issue(4)：245-256,12.DOI:10.3779/j.issn.1009-3419.2024.101.05
COX-2/PGE2/EP4轴诱导巨噬细胞功能活化在NSCLC发展过程中的作用
Role of COX-2/PGE2/EP4 Axis-induced Macrophage Functional Activation in NSCLC Development
摘要
Abstract
Background and objective Tumor microenvironment(TME)is one of the important factors in tu-morigenesis and progression,in which tumor-associated macrophages(TAMs)play an important role in non-small cell lung cancer(NSCLC)progression.However,the mechanism of TAMs in NSCLC progression remains unclear,so this study aimed to investigate the role of TAMs in NSCLC progression and to find potential therapeutic targets.Methods Gene Expression Profiling Interactive Analysis(GEPIA)database was used to analyze the expression of prostaglandin E2 receptor 4(EP4)mRNA in NSCLC and normal lung tissues;the protein expression levels of cyclooxygenase-2(COX-2),EP4,cluster of differentiation 86(CD86),CD163 and CD31 were detected by immunohistochemistry(IHC)in 120 NSCLC tissues and 24 paracancerous tissues specimens.The nude mouse lung adenocarcinoma cell A549 and macrophage RAW264.7 co-transplanted tumor model was established.And the samples were collected by gavage with EP4 inhibitor E7046,and then stained with hematoxylin-eosin(HE),IHC,and immunofluorescence(IF),and then detected by Western blot for the epithelial mesenchymal transformation(EMT)of the tumor tissues of the nude mice in each group.Western blot was used to detect the expressions of EMT related protiens in each group of nude mice;full-length transcriptome sequencing was used to screen the key genes causing liver me-tastasis and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis was performed.Results EP4 mRNA expression level in NSCLC tissues was generally lower than that in normal lung tissues(P<0.05);COX-2,EP4,CD163,CD31 proteins were differentially expressed in NSCLC tissues and adjacent tissues,and differences were observed in many clinico-pathological parameters of NSCLC patients;RAW264.7 shortened the latency period of tumorigenesis of A549 and promoted the proliferation of tumors and liver metastasis of tumors,and E7046 could reduce tumor cell proliferation activity,tumor tissue vascular density and M2-type macrophage infiltration in nude mice;IF staining showed that macrophages were mainly distributed around the metastatic foci of tumors;Western blot results showed that compared with A549 alone transplantation group,the relative expression of E-cadherin protein in tumor tissues of mice in A549 and RAW264.7 co-transplantation group was significantly decreased,and the difference was statistically significant(P<0.05),while the relative expression of N-cadherin protein was up-regulated,but the difference was not statistically significant(P>0.05);the main pathways enriched in the differ-ential genes of the full-length transcriptome were the PI3K-AKT and MAPK signaling pathways.Conclusion During NSCLC development,the COX-2/PGE2/EP4 axis may promote tumor progression by inducing macrophage functional activation,and EP4 may be a potential new target for tumor immunotherapy.This study provides new perspectives and ideas for in-depth exploration of the mechanisms of NSCLC development,as well as a theoretical basis for the development of new therapeutic strategies for NSCLC.关键词
肺肿瘤/PGE2/EP4/巨噬细胞/肿瘤免疫Key words
Lung neoplasms/PGE2/EP4/Macrophage/Tumor immunity引用本文复制引用
赵娟,朱倩莹,张宇,黎贵芸,张映林,李方方,边莉..COX-2/PGE2/EP4轴诱导巨噬细胞功能活化在NSCLC发展过程中的作用[J].中国肺癌杂志,2024,27(4):245-256,12.基金项目
本研究受国家自然科学基金项目(No.82360523、No.82060423)、昆明医科大学肺部恶性肿瘤精准病理诊断科技创新团队(No.CXTD202210)和兴滇人才计划"名医专项"(No.RLMY20220018)资助 This study was supported by the grants from National Natural Science Foundation of China(No.82360523,No.82060423),Science and Technology Innovation Team for Precision Pathological Diagnosis of Lung Malignant Tumours of Kunming Medical University(No.CXTD202210)and"Famous Doctor Special Project"of Xingdian Talent Programme(No.RLMY20220018)(All to Li BIAN). （No.82360523、No.82060423）
