MicroRNA-431-5p Inhibits the Tumorigenesis of Osteosarcoma Through Targeting PANX3
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Updated: Dec 7, 2025

An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
Shengliang Sun1, Lei Fu1, Gen Wang1
1Department of Orthopedic Trauma, The 89th Army Hospital of the Chinese People's Liberation Army, Weifang, Shandong, People's Republic of China.
Purpose:
This study aimed to evaluate the regulatory role of miR-431-5p on the tumorigenesis of osteosarcoma (OS) and the underlying mechanism involving pannexin 3 (PANX3).
Methods:
qRT-PCR was applied to measure the expression of miR-431-5p in OS tissues and cells. PANX3 and miR-431-5p were overexpressed in U2OS and HOS cells. The cell viability and apoptosis were determined by MTT and FITC/PI double staining assay, respectively. Transwell assay was performed to detect cell migration and invasion. The protein expression of cleave-caspase-3 and MMP-2/-9 was detected by Western blot. The target relationship between miR-431-5p and PANX3 was predicated by ENCORI and identified by DLR assay. The anti-tumor effect of miR-431-5p was further analyzed in a xenograft tumor model in mice.
Results:
MiR-431-5p expression was down-regulated in OS tissues and negatively correlated with lymph node metastasis and TNM stage. Over-expression of miR-431-5p induced cell apoptosis, inhibited cell proliferation, migration and invasion, up-regulated cleave-caspase-3, and down-regulated MMP-2 and -9 in OS cells. Over-expression of miR-431-5p also inhibited the growth of tumor xenografts in mice. In addition, PANX3 was a target of miR-431-5p. Over-expression of PANX3 reversed the anti-tumor effect of miR-431-5p mimics on U2OS and HOS cells.
Conclusion:
Up-regulation of miR-431-5p suppressed the tumorigenesis of OS via targeting PANX3.
Insights
MicroRNA-431-5p (miR-431-5p) suppresses osteosarcoma (OS) growth by targeting pannexin 3 (PANX3). Upregulating miR-431-5p inhibits OS cell proliferation, migration, and invasion, offering a potential therapeutic strategy.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Osteosarcoma (OS) is a primary bone malignancy with significant metastatic potential.
- MicroRNAs (miRNAs) play crucial roles in cancer development and progression.
- Dysregulation of specific miRNAs, like miR-431-5p, is implicated in OS tumorigenesis.
Purpose of the Study:
- To investigate the regulatory function of miR-431-5p in osteosarcoma.
- To elucidate the mechanism by which miR-431-5p affects OS, focusing on its interaction with pannexin 3 (PANX3).
Main Methods:
- Quantitative real-time PCR (qRT-PCR) to assess miR-431-5p expression in OS tissues and cells.
- In vitro assays (MTT, Transwell) to evaluate cell viability, apoptosis, migration, and invasion following miR-431-5p and PANX3 manipulation.
- Western blot analysis for key proteins (cleave-caspase-3, MMP-2/-9).
- Dual-luciferase reporter (DLR) assay to confirm the direct targeting of PANX3 by miR-431-5p.
- In vivo xenograft mouse model to assess the anti-tumor efficacy of miR-431-5p.
Main Results:
- miR-431-5p was significantly downregulated in OS tissues and correlated with advanced tumor stage and lymph node metastasis.
- Overexpression of miR-431-5p inhibited OS cell proliferation, migration, and invasion, while promoting apoptosis.
- miR-431-5p overexpression led to increased cleave-caspase-3 and decreased MMP-2/-9 levels.
- PANX3 was confirmed as a direct target of miR-431-5p, and its overexpression reversed the anti-tumor effects of miR-431-5p.
- In vivo studies demonstrated that miR-431-5p suppressed tumor growth in a xenograft model.
Conclusions:
- miR-431-5p acts as a tumor suppressor in osteosarcoma.
- The anti-tumorigenic effects of miR-431-5p are mediated through the inhibition of its target gene, PANX3.
- miR-431-5p represents a potential therapeutic target for osteosarcoma treatment.
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