二甲双胍下调MCP1促进巨噬细胞抑制结肠癌肝转移  

Metformin Enhances the Antitumor Effects of Macrophages in Colorectal Cancer Liver Metastasis by Down-regulatingMCP1

作　　者：刘永凡 魏昇 郭子成 崔健 朱驿 周大臣[1] 耿小平[1] 崔笑[1] LIU Yong-fan;WEI Sheng;GUO Zi-cheng;无(Dept of General Surgery,The Second Affiliated Hospital of Anhui Medical University,Anhui,Hefei 230601;The First Clinical Medical College of Anhui Medical University,Heifei 230032)

机构地区：[1]安徽医科大学第二附属医院普外科,合肥230601 [2]安徽医科大学第一临床医学院,合肥230032

出　　处：《肝胆外科杂志》2025年第1期66-72,共7页Journal of Hepatobiliary Surgery

基　　金：西藏自治区自然科学基金(基金编号:XZ202401ZR0021);安徽省高校科研项目计划(基金编号:2024AH050797);安徽省高等学校省级质量工程项目(基金编号:JYXM1140)。

摘　　要：目的研究二甲双胍通过单核细胞趋化蛋白1(monocyte chemoattractant proteinl,MCP1)调控巨噬细胞对结肠癌肝转移(LMCC,liver metastasis of colorectal cancer)的影响。方法通过构建巨噬细胞-结肠癌细胞共培养模型,诱导肿瘤相关巨噬细胞。脾脏注射切脾法构建小鼠结肠癌肝转移模型。采用免疫组织化学染色检测结肠癌肝转移组织中肿瘤相关巨噬细胞(tumor associated macrophage,TAM)标记物CD206和MCP1表达。通过细胞克隆实验和细胞Transwell实验检测结肠癌细胞增殖、迁移和侵袭能力;酶联免疫吸附实验检测(ELASA,Enzyme-linked immunosorbent assay)检测细胞培养液中MCP1浓度。结果CD206和MCP1在结肠癌肝转移组织表达较非癌肝组织增高。在体外肿瘤与巨噬细胞共培养模型中,MCP1表达升高,巨噬细胞诱导极化为CD206高表达型TAMs,肿瘤细胞增殖、迁移和侵袭能力增强。在共培养模型中使用二甲双胍后,肿瘤细胞AMPK磷酸化水平增高及磷酸化NF-κB水平降低,MCP1表达水平下降。在小鼠模型中,同样表现出MCP1表达与结肠癌肝转移中TAM数量和肿瘤大小正性相关,二甲双胍降低MCP1在肿瘤中的表达水平和TAMs在肿瘤中的数量,抑制肿瘤在小鼠体内的生长。结论二甲双胍通过下调MCP1表达促进巨噬细胞对结肠癌肝转移肿瘤免疫作用。1 Objective To investigate the function of MCPI in regulating macrophages in the progression of LMCC.Methods A co-culture model of macrophages and colon cancer cells was constructed to induce TAM.A mouse model of LMCC was established using the splenic injection and splenectomy method.TAMs were induced by constructing a macrophage-colorectal cancer cell co-culture model.The expression of CD206 and MCP1 in LMCC was detected by immunohistochemical staining.Colony formation assay and tran-swell assay were used to detect the proliferation,migration and invasion ability of colorectal cancer cells.ELASA assay was used to measure the concentration of MCP1 in the cell culture medium.Results CD206 and MCP1 were highly expressed in LMCC tumor tissue in comparison of adjacent non-tumor tissue.In the co-culture model,macrophages were polarized into TAMs,and the expression of MCPl in the culture medium was found increased.The capacity of proliferation,migration and invasion of tumor cells were enhanced.Metformin treatment elevated AMPK phosphorylation levels and reduced NF-κB phosphorylation.In vivo studies revealed that MCP1 in-creased TAM numbers and accelerated tumor progression.Metformin inhibited tumor growth and diminished TAM numbers via MCPI modulation.Conclusion Metformin enhanced the antitumor effects of macrophages in LMCC through MCP1.

关 键 词：结肠癌肝转移 单核细胞趋化蛋白1 二甲双胍 肿瘤相关巨噬细胞 上皮-间质转化 

分 类 号：R735.7[医药卫生—肿瘤]