乳腺癌来源的琥珀酸促进巨噬细胞极化的机制  

Mechanism of macrophage polarization induced by succinic acid derived from breast cancer

作　　者：朱会利[1] 王清云 林婷[3] ZHU Huili;WANG Qingyun;LIN Ting(Department of General Surgery,Handan Hospital of Integrated Traditional Chinese and Western Medicine,Handan 056001;Laboratory Department,The First Affiliated Hospital of Xingtai Medical College,Xingtai 054001;Department of Breast Surgery,Tangshan People′s Hospital,Tangshan 063001,China)

机构地区：[1]邯郸市中西医结合医院普外科,河北邯郸056001 [2]邢台医学高等专科学校第一附属医院检验科,河北邢台054001 [3]唐山市人民医院乳腺外科,河北唐山063001

出　　处：《生物技术》2024年第4期504-510,496,共8页Biotechnology

基　　金：河北省医学科学研究课题(202111341);邢台市重点研发计划项目(2020ZC198)。

摘　　要：[目的]探讨乳腺癌细胞分泌琥珀酸盐促进巨噬细胞极化的潜在机制。[方法]不同乳腺癌细胞系或正常乳腺上皮细胞的培养上清培养人巨噬细胞后,分析TAM标志物Arg1、Fizz1、Mgl1的表达以及巨噬细胞极化为TAM水平。检测乳腺癌细胞和正常乳腺上皮细胞中琥珀酸盐的水平并评估其对巨噬细胞极化为TAM的影响。分子对接和表面等离子共振分析琥珀酸与琥珀酸受体(SUCNR1)的结合位点。[结果]乳腺癌细胞上清培养的巨噬细胞中TAM标志物的表达水平上升(1.00±0.08 vs 19.71±1.80,P<0.05),并且巨噬细胞极化为TAM的水平上升(1.00±0.10 vs 4.89±0.44,P<0.05)。乳腺癌细胞上清中琥珀酸的水平显著上升[(0.09±0.01 vs 0.63±0.04)mmol/L,P<0.05]。不同浓度琥珀酸处理后,巨噬细胞中TAM标志物的表达水平呈浓度依赖性上升(1.00±0.01 vs 26.14±0.21,P<0.05),并且巨噬细胞极化为TAM的水平上升(1.00±0.04 vs 4.10±0.23,P<0.05)分子对接和表面等离子共振显示琥珀酸盐与SUCNR1结合于R81、N173和S178位点。[结论]乳腺癌细胞分泌的琥珀酸盐与巨噬细胞SUCNR1的R81、N173和S178结合后促进了巨噬细胞发生M2极化为TAM(1.00±0.10 vs 4.89±0.44)。[Objective]To explore the potential mechanism of breast cancer cells secreting succinate to promote the polarization of macrophages.[Method] After cultured human macrophages with different breast cancer cell lines or normal breast epithelial cells, the expression of TAM markers Arg1,Fizz1 and Mgl1 and the normalization of macrophages to TAM level were analyzed.To detect the level of succinate in breast cancer cells and normal breast epithelial cells and evaluate its effect on the polarization of macrophages into TAM.Molecular docking and surface plasmon resonance analysis of the binding site of succinic acid to succinic acid receptor(SUCNR1).[Result] The expression level of TAM markers in macrophages cultured with supernatant of breast cancer cells was increased(1.00±0.08 vs 19.71±1.80,P<0.05),and the level of TAM polarization in macrophages was increased(1.00±0.10 vs 4.89±0.44,P<0.05).The level of succinic acid in the supernatant of breast cancer cells was significantly increased [(0.09±0.01 vs 0.63±0.04)mmol/L,P<0.05].After treatment with succinic acid at different concentrations, the expression level of TAM markers in macrophages increased in a concentration-dependent manner(1.00±0.01 vs 26.14±0.21,P<0.05).Moreover, the level of macrophage polarization into TAM increased(1.00±0.04 vs 4.10±0.23,P<0.05).Molecular docking and surface plasmon resonance showed that succinate and SUCNR1 bind to R81,N173,and S178 sites.[Conclusion]The binding of succinate secreted by breast cancer cells to R81,N173 and S178 of macrophage SUCNR1 promoted the polarization of macrophage M2 to TAM(1.00±0.10 vs 4.89±0.44).

关 键 词：乳腺癌 琥珀酸 巨噬细胞极化 SUCNR1 Arg1 FIZZ1 Mgl1 分子对接 

分 类 号：Q784[生物学—分子生物学]