Identification and Characterization of MicroRNAs Associated with Borax-mediated Anti-tumor Activity through High-throughput Technology.

The primary aim of this study was to explore the involvement of miRNAs in the anti-tumor effects of borax through high-throughput sequencing analysis. Total RNA was extracted and purified from HepG2 cells treated with 4 mM borax for 2 or 24 hours. The samples were subjected to microarray analysis using a human miRNA array. Differentially expressed miRNAs were identified through a volcano plot and heatmap analysis and validated using quantitative PCR. A proteinprotein interaction (PPI) network was constructed, and hub genes were identified using Cytoscape software. Exposure to borax significantly altered miRNA expression levels in HepG2 cells. After 2 or 24 hours of borax treatment, 14 miRNAs were upregulated, respectively, while 3 miRNAs were downregulated compared to the control group (≥2- fold change, P<0.05). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses revealed that the target genes of differentially expressed miRNAs were primarily involved in the MAPK, TGF-β, and NFκB signaling pathways in the 2-hour treatment group. In contrast, the 24-hour treatment group showed involvement in the Ras signaling pathway, the forkhead box O signaling pathway, and cellular senescence. PPI network analysis identified NACC2, CACNB1, and FZD6 as hub genes in the 2-hour treatment group, while CDK6, BCL-2, IGF1R, BTG2, AGO2, and DLGAP3 were identified as hub genes in the 24-hour treatment group. The findings suggested that the anti-tumor effects of borax may be associated with changes in miRNA expression. This study established a potential miRNA-mRNA regulatory network related to tumor biology, providing a comprehensive understanding of the molecular mechanisms and offering new therapeutic targets for liver cancer.