蟾毒灵抑制M2型巨噬细胞介导的结直肠癌迁移和上皮间质转化  

Inhibition of M2-type macrophage-mediated migration and epithelial mesenchymal transition in colorectal cancer by bufalin

作　　者：唐东豪 陈进宝 贾琳琳 沈东晓 尚靖 冯月娇 卢佳豪 肖增友 何钰洁 王杰[1,2,3] Tang Donghao;Chen Jinbao;Jia Linlin;Shen Dongxiao;Shang Jing;Feng Yuejiao;Lu Jiahao;Xiao Zengyou;He Yujie;Wang Jie(Dept of General Surgery,Shanghai Putuo Central School of Clinical Medicine,Anhui Medical University,Shanghai 200062;The Fifth School of Clinical Medicine,Anhui Medical University,Hefei 230032;Dept of General Surgery,Putuo Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200062)

机构地区：[1]安徽医科大学上海普陀中心临床学院普外科,上海200062 [2]安徽医科大学第五临床医学院,合肥230032 [3]上海中医药大学附属普陀医院普外科,上海200062

出　　处：《安徽医科大学学报》2024年第2期310-315,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金面上项目(编号:81973625);上海市启明星项目(编号:22YF1441400);安徽医科大学研究生科研与实践创新项目资助(编号:YJS20230098);上海市中医医院第三届中医传承人才项目(编号:2023zycc02)。

摘　　要：目的探讨蟾毒灵(BU)抑制M2型巨噬细胞介导结直肠癌转移的作用。方法用佛波酯(PMA)诱导人急性白血病单核细胞(THP-1)分化为M0型巨噬细胞,48h后用含有白细胞介素-4(IL-4)和IL-13的培养基处理M0型巨噬细胞,通过酶联免疫吸附试验(ELISA)、形态学、实时荧光定量(RT-qPCR)实验观察其表面标志物和形态变化,RT-PCR和ELISA实验检测M2型巨噬细胞的表面标志物转化生长因子-β(TGF-β)、IL-10。通过ELISA实验比较M2型巨噬细胞和结直肠癌细胞HCT116上清液中IL-6分泌水平,通过Transwell实验、划痕实验、RT-qPCR和Western blot实验检测条件培养基对结直肠癌细胞HCT116的的影响。在条件培养基中加入BU后,通过Western blot、Transwell实验、划痕实验、和RT-qPCR实验观察可以HCT116中AKT/PI3K蛋白以及迁移和上皮间质转化能力的变化。结果将THP-1成功诱导成为M2型巨噬细胞。M2型巨噬细胞通过分泌IL-6激活了HCT116中AKT/PI3K蛋白磷酸化,促进了其迁移和上皮间质转化能力。BU可以抑制M2巨噬细胞介导的HCT116迁移和上皮间质转化。结论M2型巨噬细胞释放IL-6激活了结直肠癌细胞AKT/PI3K信号通路,促进了其迁移和上皮间质化。此外,BU可以抑制其促迁移和上皮间质化作用。Objective To investigate the role of bufalin(BU) in inhibiting M2-type macrophage-mediated colorectal cancer metastasis.Methods Human acute leukemia mononuclear cells(THP-1) were differentiated into M0 macrophages using phorbol ester induction(PMA) for 48 hours.The M0 macrophages were then treated with IL-4 and IL-13 medium.Surface markers and morphological changes were observed through ELISA,morphology,and RT-qPCR experiments.RT-PCR and ELISA experiments were conducted to detect the surface markers TGF-β and IL-10 of M2 macrophages.The secretion level of IL-6 in the supernatant of M2 macrophages and colorectal cancer cells HCT116 was compared using ELISA.Additionally,the effect of conditioned medium on colorectal cancer cell HCT116 was assessed through Transwell,Wound healing,RT-qPCR,and Western blot experiments.Subsequently,bufalin was added to the conditioned medium and the changes in AKT/PI3K protein,migration,and epithelial-mesenchymal transition ability in HCT116 were observed using Western blot,Transwell,Wound healing and RT-qPCR experiments.Results THP-1 were successfully differentiated into M2 macrophages.The activation of AKT/PI3K protein in HCT116 cells was induced by the secretion of IL-6 from M2 macrophages,which in turn promoted the migration and epithelial-mesenchymal transition ability of the HCT116 cells.The migration and epithelial-mesenchymal transition mediated by M2 macrophages in HCT116 cells were effectively inhibited by Bufalin.Conclusion The release of IL-6 from M2 macrophages activates the AKT/PI3K signaling pathway in colorectal cancer cells,thereby promoting their migration and epithelial-mesenchymal transition capacity.Moreover,bufalin exhibits inhibitory effects on this effect.

关 键 词：蟾毒灵 M2型巨噬细胞 结直肠癌 

分 类 号：R735.3[医药卫生—肿瘤]