β-Sitosterol blocks the LEF-1-mediated Wnt/β-catenin pathway to inhibit proliferation of human colon cancer cells

• Published in: Cellular signalling 2023-04, Vol.104, p.110585-110585, Article 110585
• Main Authors: Gu, Shengliang, Liu, Fahui, Xie, Xueheng, Ding, Meng, Wang, Zhen, Xing, Xiaoyan, Xiao, Tianbao, Sun, Xiaobo
• Format: Article
• Language: English
• Subjects: beta Catenin - metabolism; Bioinformatic analysis; catenin pathway; Cell and Molecular Biology; Cell Biology; Cell Line, Tumor; Cell Proliferation; Cell- och molekylärbiologi; Colon cancer; Colonic Neoplasms - pathology; epithelial-mesenchymal transition; expression; Gene Expression Regulation, Neoplastic; Humans; LEF-1; Lymphoid Enhancer-Binding Factor 1 - metabolism; mutations; Proliferation; Sitosterol; statistics; target; Wnt; Wnt Signaling Pathway; Wnt/β-catenin pathway; β-Sitosterol
• ISSN: 0898-6568; 1873-3913
• DOI: 10.1016/j.cellsig.2022.110585

This study aimed to investigate the LEF-1-mediated Wnt/β-catenin pathway for its biological functions and prognostic value in colon cancer (CC). Furthermore, the potential molecular mechanism of β-sitosterol in CC was investigated in vitro. Clinical information and gene expression profiles from CC patients were obtained based on Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) databases. In addition, we applied R software “Limma” package for the differential analysis of LEF-1 between cancer and para-carcinoma tissue samples. Kaplan–Meier (KM) survival analysis was adopted for analyzing whether LEF-1 was of prognostic significance. Moreover, gene set enrichment analysis (GSEA) was adopted for pathway enrichment analysis and visualization. In addition, CCK8, plate cloning, scratch and high-content screening (HCS) imaging assays were performed to examine the therapeutic efficacy of β-sitosterol in human CC HCT116 cells. siRNA technology was employed to knock down LEF1 expression in HCT116 cells. qRT-PCR and Western-blot (WB) analysis were carried out to analyze the HCT-116 mRNA and protein expression levels, respectively. LEF-1 was up-regulated within CC and acted as an oncogenic gene. LEF-1 up-regulation predicted the dismal prognostic outcome and activated the Wnt/β-catenin pathway. β-sitosterol effectively suppressed HCT116 cells proliferation and invasion. For the mechanism underlying β-sitosterol, β-sitosterol was found to significantly down-regulate LEF-1 gene and protein expression and disrupt Wnt/β-catenin pathway transmission in HCT116 cells. After suppressing LEF-1 expression, its downstream targets including C-myc, Survivin and CCND1 were also down-regulated. According to our results, LEF-1 down-regulation can effectively block Wnt/β-catenin pathway, inhibit CC cell growth and migration. Collectively, β-sitosterol can be used to treat CC, which can provide anti-tumor activity by targeting LEF-1. •LEF-1 gene is highly expressed in colon cancer, which is associated with poor prognosis•The expression of LEF-1 was significantly up-regulated in the high TNM stage compared with the low TNM stage•LEF-1 can reverse regulate the expression of genes in the Wnt/β-Catenin pathway•β-sitosterol can inhibit the proliferation of colon cancer cells by down-regulating the expression of LEF-1