Formononetin inhibits immune escape of esophageal cancer cells through the ERK signaling pathway
ZHENG Shizhen
WANG Weichen
MA Huanghuang
Abstract：Objective To explore the mechanism of Formononetin in mediating immune escape in esophageal cancer cells through the extracellular signal-regulated kinase(ERK)signaling pathway.Methods The esophageal cancer TE-1 cell line was used for primary culture.The third-generation(P3)monolayer TE-1 cells were induced with Formononetin at varying concentrations(0,50,100,200μg/mL)for 24 hours.Proliferation,migration,and invasion abilities of TE-1 cells were detected through MTT assay,wound healing assay and Transwell assay,respectively.Relative contents of immune escape-related factors(interleukin-4[IL-4],IL-10,tumour necrosis factor alpha[TNF-α])in cell supernatant were measured by enzyme-linked immunosorbent assay(ELISA).Western blot was used to detect the protein expressions of the key immune escape factor B7 homolog 1(B7-H1)and ERK signaling pathway associated proteins(ERK1/2,c-Fos,c-Jun).Esophageal cancer TE-1 cells were then induced with blank control,Formononetin,the ERK signaling pathway inhibitor SCH772984,and Formononetin+SCH772984.Expression levels of key ERK signaling pathway associated proteins(ERK1/2,c-Fos,c-Jun)were detected to determine the targeted regulation of the ERK signaling pathway by Formononetin.Results With the prolongation of cell culture,TE-1 cells induced with blank control,Formononetin,SCH772984,and Formononetin+SCH772984 all presented significantly increased proliferation(P＜0.05).Compared with those induced with blank control,Formononetin treatment significantly downregulated the proliferation,migration,and invasion abilities of esophageal cancer cells in a dose-dependent manner(P＜0.05).Formononetin treatment significantly downregulated the immune co-stimulatory factor B7-H1 and immune escape-related factors(IL-4,IL-10,TNF-α)in a dose-dependent manner(P＜0.05).Formononetin treatment significantly downregulated protein expressions of key proteins in the ERK signaling pathway(ERK1/2,c-Fos,c-Jun)in a dose-dependent manner(P＜0.05).Compared with those induced with blank control,treatment of either Formononetin or SCH772984 significantly downregulated the protein expressions of key ERK signaling pathway associated proteins(ERK1/2,c-Fos,c-Jun)(P＜0.05).The treatment of Formononetin+SCH772984 further reduced the activity of the ERK signaling pathway,and protein expressions of ERK1/2,c-Fos,and c-Jun(P＜0.05).Conclusion Formononetin reduces the immune escape ability,cell proliferation,migration,and invasion of esophageal cancer cells by inhibiting the ERK signaling pathway,and ultimately inhibits the distant metastases.It is worth further clinical research.
Keywords：Formononetinesophageal cancerextracellular signal-regulated kinase(ERK)signaling pathwayimmune escapemalignant progression
Publication Date：2024-10-25
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 3045-3050 )