The effect of highly metastatic colorectal cancer cells on immune escape of colorectal cancer by secreting KLF4-induced M2 macrophages polarization
- Author:
Jing Shang
1
;
Chang Lu
2
;
Yangbo Hou
3
;
Qin Yuan
2
;
Wei Li
2
;
Haijing Wang
4
;
Jinbao Chen
5
Author Information
1. Dept of Traditional Chinese Medicine Oncology , Putuo Hospital , Shanghai University of Traditional Chinese Medicine , Shanghai 200062; Dept of Radiology , Putuo Hospital , Shanghai University of Traditional Chinese Medicine , Shanghai 200062
2. Dept of General Surgery , Putuo Hospital , Shanghai University of Traditional Chinese Medicine , Shanghai 200062
3. Dept of Neurology , Putuo Hospital , Shanghai University of Traditional Chinese Medicine , Shanghai 200062
4. Dept of Pharmacy , Putuo Hospital , Shanghai University of Traditional Chinese Medicine , Shanghai 200062
5. Dept of Traditional Chinese Medicine Oncology , Putuo Hospital , Shanghai University of Traditional Chinese Medicine , Shanghai 200062
- Publication Type:Journal Article
- Keywords:
highly metastatic colorectal cancer cells;
KLF4;
tumor-associated macrophages;
immune escape;
PD-L1
- From:
Acta Universitatis Medicinalis Anhui
2025;60(6):1036-1042
- CountryChina
- Language:Chinese
-
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-culture 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-Mφ and 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 expression of M2 macrophage markers interleukin(IL-10) and transforming growth factor-β(TGF-β) were higher. Compared 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 decreased.
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.
- Full text:2026030216145426720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_尚靖.pdf
