高转移性结直肠癌细胞通过分泌KLF4诱导M2型巨噬细胞极化对结直肠癌免疫逃逸的影响
The effect of highly metastatic colorectal cancer cells on immune escape of colorectal cancer by secreting KLF4-induced M2 macrophages polarization
摘要
Abstract
Objective To investigate the effect and mechanism of M2 macrophage polarization induced by HCT116 with highly metastatic colorectal cancer cells on immune escape in colorectal cancer.Methods After the co-cul-ture of colorectal cancer cells conditioned medium(CM)and PMA-induced M0 macrophages,the polarization of M2 macrophages was observed by flow cytometry,real-time polymerase chain reaction(qPCR)and enzyme-linked immunosorbent assay(ELISA)experiments.The CMM0,CMSW480-Mφand CMHCT116-Mφ were co-cultured with HCT116 cells,and the expression of programmed death-ligand 1(PD-L1)was detected by Western blot and qPCR.At the same time,the stimulated HCT116 cells were co-cultured with human T lymphocytes to detect the survival of HCT116 cells and the levels of tumor necrosis factor-α(TNF-α)and interferon-γ(IFN-γ).The difference of kruüppel-like factor 4(KLF4)expression between SW480 and HCT116 cells was detected by Western blots,qPCR and ELISA.After pretreatment of HCT116 cells with KFL4 inhibitor Kenpaullone(Ken),the CMHCT116 and CMHCT116+Ken were co-cultured with M0 macrophages and the polarization of M2 macrophages was observed by flow cytometry,qPCR and ELISA.The CMHCT116-Mφ and CM(HCT116+Ken)-Mφ was co-cultured with HCT116 cells,and the PD-L1 expression of HCT116 cells was detected by Western blot and qPCR.Results After the stimulation of M0 macrophages with CM,the proportion of CD11b+CD206+cells in HCT116-Mφ cells was higher,and the expres-sion of M2 macrophage markers interleukin(IL-10)and transforming growth factor-β(TGF-β)were higher.Com-pared with the CMSW480-Mφ group,the PD-L1 protein expression level was higher in the CMHCT116-Mφ group.After co-culture with T lymphocytes,the cell survival rate are the most in CMHCT116-Mφ group,while the levels of TNF-α and IFN-γ were the lowest.After the addition of Ken,the polarization ratio and markers of M2 macrophages decreased.Compared with CMHCT116-Mφ group,the expression of PD-L1 in HCT116 cells of the CM(HCT116+Ken)-Mφ group de-creased.Conclusion Highly metastatic colorectal cancer cells induce polarization of M2 macrophages by secreting KLF4,promote PD-L1 expression in colorectal cancer cells,facilitate tumor immune escape,and provide potential targets for clinical immunotherapy.关键词
高转移性结直肠癌细胞/KLF4/肿瘤相关巨噬细胞/免疫逃逸/PD-L1Key words
highly metastatic colorectal cancer cells/KLF4/tumor-associated macrophages/immune escape/PD-L1分类
临床医学引用本文复制引用
尚靖,路畅,侯阳波,袁沁,李炜,王海静,陈进宝..高转移性结直肠癌细胞通过分泌KLF4诱导M2型巨噬细胞极化对结直肠癌免疫逃逸的影响[J].安徽医科大学学报,2025,60(6):1036-1042,7.基金项目
上海市启明星基金(扬帆专项)(编号:22YF1441400) （扬帆专项）
上海市普陀区卫生健康系统科技创新项目(编号:ptkwws202409) （编号:ptkwws202409）
上海市第六人民医院联合共建科学研究基金(编号:23-LY-03) （编号:23-LY-03）
上海中医药大学附属普陀医院百人计划(编号:2022-RCLH-03) （编号:2022-RCLH-03）
上海中医药大学预算内项目(编号:2021LK055) （编号:2021LK055）
上海市卫生健康系统重点学科计划(编号:2024ZDXK0046) Shanghai Rising-Star Program(Sailing Special Project)(No.22YF1441400) （编号:2024ZDXK0046）
Technology Inno-vation Project of Shanghai Putuo District Health System(No.ptkwws202409) （No.ptkwws202409）
Shanghai Sixth People's Hospital Joint Constructed Scientific Research Fund(No.23-LY-03) （No.23-LY-03）
"100 Talents Project"of Putuo Hospital,Shanghai University of Traditional Chinese Medicine(No.2022-RCLH-03) （No.2022-RCLH-03）
Budgeted Project of Shanghai University of Tra-ditional Chinese Medicine(No.2021LK055) （No.2021LK055）
Key Discipline Plan of Shanghai Health System(No.2024ZDXK0046) （No.2024ZDXK0046）
