基于EGF-EGFR信号通路探讨阳和汤阻抑M2型肿瘤相关巨噬细胞促进乳腺癌细胞迁移侵袭的机制

Mechanism of Yanghe decoction inhibiting M2-type TAMs to promote migration invasion of triple-negative breast cancer cells based on EGF-EGFR signaling pathway

目的基于旁分泌表皮生长因子(epidermal growth factor,EGF)/表皮生长因子受体(epidermal growth factor receptor,EGFR)信号通路,探究阳和汤含药血清对M2型肿瘤相关巨噬细胞(tumor-associated macrophages,TAMs)与乳腺癌MCF-7细胞共培养体系内MCF-7细胞迁移侵袭的机制研究。方法选取人单核细胞系THP-1,体外经佛波酯(PMA)和重组人巨噬细胞群刺激因子(MCSF)诱导获得M2型TAMs模型。将MCF-7细胞与M2型TAMs采用Transwell非接触式共培养,评估其对迁移、侵袭能力的影响。随后,使用含阳和汤的药血清干预细胞,采用CCK-8法检测MCF-7细胞增殖情况,并通过划痕实验检测其横向迁移能力。利用Transwell实验分别检测细胞的侵袭和纵向迁移能力,通过ELISA法测定EGF浓度。采用Western blot法检测EGFR、MCP-1和MMP-9蛋白的表达。结果与空白组相比,阳和汤含药血清抑制了共培养前后MCF-7细胞的增殖。阳和汤含药血清对共培养前后的划痕愈合能力降低,并降低了其迁移和侵袭能力。阳和汤含药血清降低了共培养前后EGF的含量并且降低了共培养前后EGFR、MCP-1和MMP9蛋白的表达。结论阳和汤含药血清可以抑制与M2 TAMs共培养前后乳腺癌MCF-7细胞的迁移、侵袭。这一效应可能与EGF-EGFR信号通路受到抑制有关。

Aim To explore the mechanism of Yanghe decoction-containing serum on the migration and invasion of MCF-7 breast cancer cells in a co-culture system with M2 tumor-associated macrophages(TAMs)based on the paracrine epidermal growth factor(EGF)/epidermal growth factor receptor(EGFR)signaling pathway.Methods The M2-type TAMs model was induced from THP-1 monocytic cell line through in vitro treatment with phorbol 12-myristate 13-acetate(PMA)and recombinant human macrophage colony-stimulating factor(M-CSF).The MCF-7 cells were co-cultured with M2-type TAMs using a Transwell non-contact co-culture system to evaluate their effects on migration and invasion.Subsequently,the cells were intervened with serum containing Yanghe decoction,and the proliferation of MCF-7 cells was detected using the CCK-8 assay,while their lateral migration ability was assessed through scratch assays.The invasion and vertical migration abilities of the cells were evaluated separately using Transwell assays,and the concentration of EGF was measured using ELISA.Finally,the expression of EGFR,MCP-1,and MMP9 proteins was detected using Western blot.Results Compared to the control group,Yanghe decoction-containing serum inhibited the proliferation of MCF-7 cells before and after co-culture.The serum reduced the scratch healing ability before and after co-culture and decreased their migration and invasion abilities.Additionally,Yanghe decoction-containing serum reduced the levels of EGF before and after co-culture and decreased the expression of EGFR,MCP-1,and MMP9 proteins before and after+co-culture.Conclusion Yanghe decoction-containing serum can inhibit the migration and invasion of breast cancer MCF-7 cells before and after co-culture with M2 TAMs.This effect may be related to the inhibition of the EGF-EGFR signaling pathway.