Dados do Resumo

Título

TRANSCRIPTOME ANALYSES REVEAL GENES RELATED TO EPIGENETICS AND THE IMMUNE SYSTEM AS POTENTIAL NEW THERAPEUTIC TARGETS FOR MEBENDAZOLE IN GASTRIC CANCER

Introdução

Gastric cancer (GC) is among the most common neoplasms worldwide and is considered the fifth most common and lethal type of cancer. Drug repositioning is a pharmaceutical strategy for discovering new therapeutic applications for a drug already approved for treating another disease. Thus, Mebendazole (MBZ) is a benzimidazole approved for clinical use, its antitumoral activity is widely discussed in the literature, including against GC cell lines. Otherwise, its molecular targets are still poorly understood.

Objetivo

Thus, this study aims to analyze and detect, through transcriptome analysis, the molecular signature of gastric adenocarcinoma cell line AGP-01 after treatment with the drug MBZ and, thus, discover new pharmacological biomarkers of interest in treating patients with GC.

Métodos

The CG cell line AGP-01 was first treated with MBZ (1 µM) for 14 hours, then mRNA extraction was performed and the transcriptome was carried out using the microarray technique (GeneChip Exon 1.0 ST Array). Results were analyzed in R software, and the overexpressed and underexpressed genes were identified, through the Gene set enrichment analysis (GSEA) the pathways related to MBZ treatment were also evaluated. The top 3 overexpressed and underexpressed genes were validated via RT-qPCR, based on in silico analyses we also verified the mRNA levels of the MBZ targets and their relation with the overall survival (OS) rate survival of patients with GC through GEPIA (http://gepia.cancer-pku.cn/) and Kaplan-Meier Plotter ((https://kmplot.com/analysis/) software, respectively.

Resultados

In the initial analysis to verify differentially expressed genes (DEGs), it was observed that 345 genes showed hyperexpression (2.41%), 721 genes showed hypoexpression (5.04%), while 13,231 genes showed no change after treatment (92.54%). The overexpressed genes identified were CCL2, IL1A, and CDKN1A, while H3C7, H3C11, and H1-5 were the top 3 underexpressed genes after treatment. Gene set enrichment analysis (GSEA) identified 8 pathways significantly overexpressed in the treated group (p<0.05 and FDR<0.25). The validation of the expression of over and underexpressed targets by RT-qPCR confirmed the transcriptome results, MBZ increased the expression of the CCL2 (p<0.001), IL1A (p<0.001), and CDKN1A (p<0.05) genes and reduced the expression of H3C7 (p<0.0001), H3C11 (p<0.0001) and H1-5 (p<0.001). Furthermore, expression analysis in silico databases demonstrated that low expression of ILI1A and high expression of H3C11 and H1-5 are associated with a reduction in the overall survival rate of patients with GC.

Conclusões

Thus, the data highly new potential targets of MBZ in GC treatment, showing that the drug MBZ alters the transcriptome of the AGP-01 cell line, mainly by modulating the expression of histone proteins and inflammatory cytokines, indicating a possible epigenetic and immunological effect on tumor cells.

Financiador do resumo

National Council for Scientific and Technological Development (CNPq), Coordination for the Improvement of Higher Education Personnel (CAPES), and Cearense Foundation for Scientific and Technological Development Support (FUNCAP).

Palavras Chave

Gene set enrichment analysis Molecular target Transcriptomic profile