High-throughput sequencing reveals crebanine inhibits colorectal cancer by modulating Tregs immune prognostic target genes

Abstract

Crebanine, an alkaloid exhibiting sedative, anti-inflammatory, and anticancer properties, remains unexplored in terms of its anticancer potential against colorectal cancer (CRC). This study aims to bridge this knowledge gap, specifically investigating whether crebanine can suppress CRC and elucidating its underlying molecular mechanism. We employed the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay, cell scratch assay, and flow cytometry to observe the effects of crebanine on the growth, migration, and apoptosis of CRC SW480 cells, respectively. High-throughput sequencing was employed to detect differentially expressed genes (DEGs) in SW480 cells treated with crebanine. Enriched pathways of these DEGs were identified through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Genes exhibiting the highest correlation in the enriched pathway were further analyzed using clinical data from The Cancer Genome Atlas Program (TCGA) public database, utilizing R software. Crebanine effectively inhibited the proliferation, migration, and invasion of SW480 cells, with concentrations of ≥15 μg/mL promoting apoptosis. Analysis revealed that the function of DEGs linked to the most enriched pathways was associated with immune infiltration by regulatory T cells (Tregs). When analyzed in conjunction with clinical data, the genes exhibiting the highest correlation in the enrichment pathway were found to be directly associated with clinical prognostic survival. Our study demonstrates that crebanine inhibits colorectal cancer by regulating prognostic target genes related to Tregs. This finding offers a novel approach for pharmacological inhibition and Tregs-targeted therapy in CRC.