026 | Graphene oxide nanosystem for antisense miRNA-21 delivery in hepatocellular carcinoma

Paola Trischitta¹, Rosamaria Pennisi¹, Maria Teresa Sciortino¹, Marta Kutwin² | ¹Dept. of Chemical, Biological, Pharmaceutical and Environmental Sciences, University of Messina, Italy; ²Dep of Nanobiotechnology, Institute of Biology, Warsaw University of Life Sciences, Warsaw, Poland.

Nanotechnology-based strategies for targeted drug delivery have attracted considerable interest, particularly in gene regulation therapies (Di Stefano, 2023). Among emerging nanocarriers, graphene oxide (GO) has shown strong potential for microRNA (miRNA)-based treatments owing to its distinctive physicochemical properties (Kutwin et al., 2023). In the present study, we developed and characterized a GO-based nanosystem for the targeted delivery of antisense miRNA-21, a key regulator of oncogenic signaling pathways (Kutwin et al., 2021). The GO–antisense miRNA-21 nanosystem was obtained through a self-assembly approach and extensively characterized by ATR-FTIR and UV–Vis spectroscopy, zeta potential measurements, and transmission electron microscopy to assess its structural features, stability, and release behavior. The biological performance of the nanosystem was evaluated both in vitro, using HepG2 hepatocellular carcinoma cells, and in vivo, in a chicken embryo xenograft tumor model. Cellular uptake and delivery efficiency were verified using confocal microscopy and flow cytometry. Gene expression analysis of inflammation- and metastasis-related markers was performed by quantitative PCR. Results demonstrated successful delivery of antisense miRNA-21, which caused a significant downregulation of IL-8 and other pro-inflammatory genes in HepG2 cells. In vivo, treatment significantly reduced tumor volume and size, with a corresponding decrease in IL-8 expression within the tumor tissues. Overall, these findings highlight the potential of graphene oxide-based nanosystems as effective and selective carriers for antisense miRNA delivery in liver cancer therapy. The combined in vitro and in vivo approach confirms their ability to modulate gene expression and inhibit tumor progression, supporting their future role in future miRNA-targeted clinical applications.