Title:miR-488-3p Represses Malignant Behaviors and Facilitates Autophagy of
Osteosarcoma Cells by Targeting Neurensin-2
Volume: 25
Issue: 10
Author(s): Chao Yun, Jincai Zhang and Morigele*
Affiliation:
- Department of Orthopedics, The Affiliated Hospital of Inner Mongolia Medical University, Hohhot, 010059, Mongolia,
China
Keywords:
Osteosarcoma, hsa-miR-488-3p, neurensin-2, autophagy, apoptosis, cell viability, migration, invasion.
Abstract:
Objectives: Osteosarcoma (OS) is a primary bone sarcoma that primarily affects children
and adolescents and poses significant challenges in terms of treatment. microRNAs (miRNAs)
have been implicated in OS cell growth and regulation. This study sought to investigate the
role of hsa-miR-488-3p in autophagy and apoptosis of OS cells.
Methods: The expression of miR-488-3p was examined in normal human osteoblasts and OS cell
lines (U2OS, Saos2, and OS 99-1) using RT-qPCR. U2OS cells were transfected with miR-488-
3p-mimic, and cell viability, apoptosis, migration, and invasion were assessed using CCK-8, flow
cytometry, and Transwell assays, respectively. Western blotting and immunofluorescence were
employed to measure apoptosis- and autophagy-related protein levels, as well as the autophagosome
marker LC3. The binding sites between miR-488-3p and neurensin-2 (NRSN2) were predicted
using online bioinformatics tools and confirmed by a dual-luciferase assay. Functional rescue
experiments were conducted by co-transfecting miR-488-3p-mimic and pcDNA3.1-NRSN2
into U2OS cells to validate the effects of the miR-488-3p/NRSN2 axis on OS cell behaviors. Additionally,
3-MA, an autophagy inhibitor, was used to investigate the relationship between miR-
488-3p/NRSN2 and cell apoptosis and autophagy.
Results: miR-488-3p was found to be downregulated in OS cell lines, and its over-expression
inhibited the viability, migration, and invasion while promoting apoptosis of U2OS cells. NRSN2
was identified as a direct target of miR-488-3p. Over-expression of NRSN2 partially counteracted
the inhibitory effects of miR-488-3p on malignant behaviors of U2OS cells. Furthermore, miR-
488-3p induced autophagy in U2OS cells through NRSN2-mediated mechanisms. The autophagy
inhibitor 3-MA partially reversed the effects of the miR-488-3p/NRSN2 axis in U2OS cells.
Conclusion: Our findings demonstrate that miR-488-3p suppresses malignant behaviors and promotes
autophagy in OS cells by targeting NRSN2. This study provides insights into the role of
miR-488-3p in OS pathogenesis and suggests its potential as a therapeutic target for OS treatment.
