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华蟾素通过调控M2型巨噬细胞极化抑制结直肠癌转移OACSTPCD

Cinobufacini inhibits the metastasis of colorectal cancer by regulating polarization of M2 macrophages

中文摘要英文摘要

目的 探讨华蟾素通过调控M2型巨噬细胞极化抑制结直肠癌转移的作用.方法 将THP-1诱导成M0型巨噬细胞,收集HCT116细胞的条件培养基,刺激M0型巨噬细胞,通过流式细胞术、RT-qPCR、ELISA实验观察M2型巨噬细胞极化状态;收集M0型巨噬细胞和HCT116-Mφ细胞的条件培养基,刺激HCT116细胞,通过划痕实验和Transwell实验观察迁移和侵袭能力;用CCK-8实验检测华蟾素对HCT116细胞活力的影响;收集HCT116和HCT116+华蟾素的条件培养基,刺激M0型巨噬细胞,通过流式细胞术、RT-qPCR、ELISA实验观察M2型巨噬细胞极化状态;收集HCT116-Mφ细胞和(HCT116+华蟾素)-Mφ细胞的条件培养基,刺激HCT116细胞,通过划痕实验和Transwell实验观察迁移和侵袭能力的改变.结果 M0型巨噬细胞在HCT116细胞的条件培养基刺激后,形态变成梭形的细胞,CD11b+CD206+细胞比例增高,M2型巨噬细胞标志物白细胞介素-10(IL-10)及转化生长因子-β(TGF-β)表达升高;HCT116细胞在HCT116-Mφ细胞的条件培养基刺激后,细胞迁移和侵袭能力明显增强;加入华蟾素之后,不仅M2型巨噬细胞极化比例降低,M2型巨噬细胞介导的促转移效应也受到抑制.结论 HCT116细胞可以诱导M2型巨噬细胞极化,而华蟾素可通过抑制M2型巨噬细胞极化,进而抑制M2型巨噬细胞介导的肿瘤转移.

Objective To investigate the effect of cinobufacini on inhibiting colorectal cancer metastasis by regula-ting the polarization of M2 macrophages.Methods THP-1 was induced into M0 type macrophages.The condi-tioned medium of HCT116 cells was collected to stimulate M0 type macrophages.The polarization of M2 type mac-rophages was observed by flow cytometry,real-time quantitative PCR and ELISA experiments.The conditioned me-dium of M0 type macrophages and HCT116-Mφ cells was collected to stimulate HCT116 cells.The ability of migra-tion and invasion was observed by wound healing assay and Transwell assay.The effect of cinobufacini on the via-bility of HCT116 cells was detected by CCK-8 assay.The conditioned medium of HCT116 and HCT116+cinobufa-cini was collected to stimulate M0 type macrophages.The polarization of M2 type macrophages was observed by flow cytometry,real-time quantitative PCR and ELISA experiments.The conditioned media of HCT116-Mφ cells and(HCT116+cinobufacini)-Mφ cells were collected to stimulate HCT116 cells.The changes of migration and inva-sion ability were observed by wound healing assay and Transwell assay.Results After stimulation of M0 type mac-rophages in HCT116 cell conditioned medium,the morphology of M0 macrophages turned into fusiform cells,the proportion of CD11b+CD206+cells increased,and the expression of M2 macrophage markers IL-10 and TGF-β in-creased.The migration and invasion ability of HCT116 cells were significantly enhanced after stimulation in the conditioned medium of HCT1 16-Mφ cells.After the addition of cinobufacini,not only the polarization proportion of M2 macrophages decreased,but also the metastatic effect mediated by M2 macrophages was inhibited.Conclusion HCT116 cells can induce the polarization of M2 macrophages,while cinobufacini can inhibit the tumor metastasis mediated by M2 macrophages by inhibiting the polarization of M2 macrophages.

尚靖;王云;陈进宝;唐东豪;贾琳琳;李炜;于宏杰

上海中医药大学附属普陀医院肿瘤科,上海 200062上海中医药大学附属普陀医院血液科,上海 200062上海中医药大学附属普陀医院普外科,上海 200062

临床医学

华蟾素结直肠癌肿瘤相关巨噬细胞转移

cinobufacinicolorectal cancertumor-associated macrophagemetastasis

《安徽医科大学学报》 2024 (002)

224-229 / 6

上海市启明星项目(扬帆专项)(编号:22YF1441400);上海市普陀区临床特色专病建设项目(编号:2020tszb03);上海中医药大学附属普陀医院百人计划(领航计划)(编号:2022-RCLH-03);成都中医药大学"杏林学者"学科人才科研提升计划(编号:YYZX2020121)

10.19405/j.cnki.issn1000-1492.2024.02.007

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