摘要
CMTM4蛋白异常表达与肿瘤发生、发展及预后相关,其在宫颈癌中的作用和机制尚不明确。本研究旨在探讨CMTM4在宫颈癌中的表达水平及其对MDSCs分化和免疫细胞功能的调节作用和分子机制。首先,利用免疫组化染色和Western印迹检测CMTM4在宫颈癌肿瘤和癌旁组织中的表达水平。结果显示,CMTM4在宫颈癌组织中的表达水平显著低于癌旁组织(P<0.01)。通过流式细胞术检测宫颈癌患者肿瘤样本中MDSCs的分化,使用CD45、CD11b和Ly6G标记M-MDSCs,使用CD45、CD11b和Ly6C标记G-MDSCs。发现宫颈癌患者血液中M-MDSCs和G-MDSCs的百分比显著高于健康志愿者组(P<0.01)。此外,流式分析发现,CMTM4高表达提高宫颈癌患者血液中CD3+CD4+T细胞和CD3+CD8+T细胞比例(P<0.01),促进Th1和Th17细胞的活化比例(P<0.05),抑制Th2和Treg细胞的活化比例(P<0.05或P<0.01)。通过流式分析肿瘤组织中CD8+PD-1+T细胞和CD8+PD-L1+T细胞的百分比。结果显示,CMTM4高表达抑制肿瘤组织中CD8+PD-1+T细胞和CD8+PD-L1+T细胞的百分比(P<0.05)。体外研究表明,CMTM4过表达的HeLa细胞显著抑制MDSCs生成(P<0.01),PD-1过表达的HeLa细胞进一步促进了MDSCs生成(P<0.05)。此外,使用ELISA检测Th1、Th2、Th17和Treg细胞因子的水平。发现CMTM4过表达的HeLa细胞可降低T细胞的IL-10分泌水平(P<0.01),而增加IFN-γ和TNF-α的分泌水平(P<0.01),其均被PD-1过表达逆转(P<0.05或P<0.01)。综上,本研究表明,CMTM4在宫颈癌中的低表达导致MDSCs的异常分化和肿瘤的免疫逃逸,CMTM4通过调节PD-1/PD-L1通路,从而影响肿瘤T细胞的活化。因此,CMTM4可能成为治疗宫颈癌的潜在靶点。
The aberrant expression of CMTM4 protein has been implicated in tumorigenesis,development,and prognosis;however,its precise role and underlying mechanism in cervical cancer remain elusive.We aimed to investigate the expression level of CMTM4 in cervical cancer and its regulatory effect and molecular mechanism on MDSCs differentiation and immune cell function.Firstly,immunohistochemical(IHC)staining and Western blot analysis were employed to assess the expression level of CMTM4 in cervical cancer tumors and adjacent tissues.The results revealed a significant decrease in the expression level of CMTM4 in cervical cancer tissues compared to adjacent tissues(P<0.01).Differentiation of MDSCs in tumor samples from cervical cancer patients was assessed using flow cytometry,with CD45,CD11b,and Ly6G used as markers for M-MDSCs,and CD45,CD11b,and Ly6C used as markers for G-MDSCs.The percentage of M-MDSCs and G-MDSCs in the blood of cervical cancer patients was significantly higher than that observed in healthy volunteers(P<0.01).Furthermore,flow cytometry analysis demonstrated that high expression of CMTM4 increased the proportion of CD3+CD4+T cells and CD3+CD8+T cells in the blood of cervical cancer patients(P<0.01),while also promoting the activation ratio of Th1 and Th17 cells(P<0.05).Conversely,it inhibited the activation ratio of Th2 and Treg cells(P<0.05 or P<0.01).The percentage of CD8+PD-1+T cells and CD8+PD-L1+T cells within tumor tissues was determined by flow cytometry analysis which showed that high expression levels of CMTM4 suppressed their presence within tumor tissues(P<0.05).The in vitro studies demonstrated that overexpression of CMTM4 in HeLa cells significantly suppressed the generation of MDSCs(P<0.01),while overexpression of PD-1 in HeLa cells further enhanced the generation of MDSCs(P<0.05).Additionally,ELISA was employed to measure the levels of Th1,Th2,Th17,and Treg cytokines.It was observed that CMTM4 overexpression in HeLa cells reduced IL-10 secretion by T cells(P<0.01),but increased IFN-γand TN
作者
王海燕
刘建辉
阿小英
王佳倞
穆庆芬
周静
杨安琪
WANG Hai-Yan;LIU Jian-Hui;A Xiao-Ying;WANG Jia-Liang;MU Qing-Fen;ZHOU Jing;YANG An-Qi(Department of Gynecology,Qinghai University Affiliated Hospital,Xining 810001,China)
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2024年第12期1742-1750,共9页
Chinese Journal of Biochemistry and Molecular Biology
基金
2023年青海省科技厅基础研究计划项目:科研基金课题(No.2023-ZJ-777)资助。
关键词
CKLF样MARVEL跨膜结构域4
宫颈癌
髓源性抑制细胞分化
免疫逃逸
CKLF like MARVEL transmembrane domain containing 4(CMTM4)
cervical cancer
myeloid-derived suppressor cells(MDSCs)differentiation
immune escape
