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

• Objectives: This study aimed to investigate the LEF-1-mediated Wnt/beta-catenin pathway for its biological functions and prognostic value in colon cancer (CC). Furthermore, the potential molecular mechanism of beta-sitosterol in CC was investigated in vitro.Methods: 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 beta-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.Results: 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/beta-catenin pathway. beta-sitosterol effectively suppressed HCT116 cells proliferation and invasion. For the mechanism underlying beta-sitosterol, beta-sitosterol was found to significantly down-regulate LEF-1 gene and protein expression and disrupt Wnt/beta-catenin pathway transmission in HCT116 cells. After suppressing LEF-1 expression, its downstream targets including C-myc, Survivin and CCND1 were also down-regulated.Conclusion: According to our results, LEF-1 down-regulation can effectively block Wnt/beta-catenin pathway, inhibit CC cell growth and migration. Collectively, beta-sitosterol can be used to treat CC, which can provide anti-tumor activity by targeting LEF-1.

Subject headings

MEDICIN OCH HÄLSOVETENSKAP  -- Medicinska och farmaceutiska grundvetenskaper -- Cell- och molekylärbiologi (hsv//swe)

MEDICAL AND HEALTH SCIENCES  -- Basic Medicine -- Cell and Molecular Biology (hsv//eng)

Keyword

Colon cancer

?-Sitosterol

LEF-1

Wnt

?-catenin pathway

Proliferation

Bioinformatic analysis

epithelial-mesenchymal transition

target

statistics

expression

mutations

Cell Biology

Publication and Content Type

ref (subject category)

art (subject category)