贞术消积汤通过FOXO信号通路抑制肝癌迁移与侵袭的机制研究OA

Objective To investigate the effect of Zhenzhu Xiaoji Decoction(贞术消积汤,ZZXJD)on the migration and invasion of liver cancer cells,focusing on the FOXO signaling pathway.Methods Wound healing assay,colony formation assay,and Transwell invasion assay were conducted to evaluate the effects of ZZXJD on the migration,invasion,and clonogenicity of liver cancer cells.The mRNA and protein expression levels of key factors in the FOXO pathway were detected by quantitative real-time PCR(qRT-PCR)and Western blot(WB),respectively.Cellular localization and expression of these proteins were observed by immunofluorescence(IF)assay.A xenograft model was established in nude mice using human liver cancer cells.The mice were randomly assigned to a control group and low/medium/high-dose ZZXJD groups.The tumor inhibition rate and organ coefficients were calculated,and HE staining was performed to observe metastatic foci in the liver and lungs.Results ZZXJD inhibited the colony formation,migration,and invasion abili-ties of liver cancer cells in a dose-dependent manner.ZZXJD treatment downregulated the mRNA and protein levels of IGF1,IGF1R,phosphorylated FOXO1(p-FOXO1),MMP2,MMP9,and VEGFA,and upregulated those of FOXO1 and FOXO6.In vivo,ZZXJD treatment increased the tumor inhibition rate and decreased the spleen organ coefficient.Moreover,the number of metastatic foci in the liver and lungs was reduced in all ZZXJD-treated groups.Conclusion ZZXJD may inhibit the migration and invasion of liver cancer cells by upregulating FOXO1 and FOXO6,and downregulating IGF1,IGF1R,p-FOXO1,MMP2,MMP9,and VEGFA at both the mRNA and protein levels.