Ulfatun Nisa, Indwiani Astuti, Ronny Martien, Dhani Rinaldi Maulana, Ysrafil Ysrafil, Leonny Dwi Rizkita,
Volume 18, Issue 1 (3-2026)
Abstract
Background: Hepatocellular carcinoma (HCC) is one of the most lethal malignancies worldwide, and dysregulation of microRNAs has been increasingly recognized as a critical driver in its pathogenesis. Among them, miR-217-5p has been reported as a tumor-suppressive miRNA, yet its molecular role and therapeutic potential in HCC remain largely unclear.
Methods: In silico analyses were performed using a network pharmacology approach to construct a protein–protein interaction (PPI) network of hepatocellular carcinoma (HCC)-associated genes targeted by miR-217-5p, followed by hub gene identification using CytoHubba. Functional enrichment analyses, including Gene Ontology (GO) and KEGG pathways, were conducted using ShinyGO, while expression and survival analyses were validated using UALCAN, GEPIA2, and the Human Protein Atlas. In vitro validation was performed by delivering miR-217-5p mimics into HepG2 cells using chitosan nanoparticles (CS-NPs), followed by qRT-PCR analysis of endogenous miRNA expression, KRAS suppression, and p53 restoration.
Results: A total of 152 potential target genes of miR-217-5p in HCC were identified, which were predominantly enriched in key biological processes related to regulation of cell population proliferation, as well as cancer-associated pathways including PI3K–Akt signaling and hepatocellular carcinoma pathways. PPI analysis, following the removal of disconnected nodes, generated a highly interconnected network comprising 146 nodes and 1042 edges, indicating extensive molecular interactions among the identified targets. Furthermore, hub gene analysis of the PPI network identified PTEN, KRAS, ESR1, HIF1A, and CDH1 as central regulatory genes. Notably, in vitro validation demonstrated that miR-217-5p delivered via chitosan nanoparticles (CS-NPs) successfully restored endogenous miR-217-5p expression and significantly suppressed KRAS expression, exhibiting a strong inverse correlation (r = −0.937, p < 0.0001). In addition, p53 protein levels were markedly increased following treatment.
Conclusion: Our findings demonstrate that miR-217-5p exerts tumor-suppressive effects in HCC by directly targeting KRAS and restoring p53 expression. Delivery of miR-217-5p using chitosan nanoparticles represents a promising miRNA replacement therapy strategy and highlights its potential as both a therapeutic agent in HCC.
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