miR-129-5p inhibits clear cell renal cell carcinoma cell proliferation, migration and invasion by targeting SPN
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Updated: Nov 5, 2025
An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
Bin Gao1, Lijuan Wang1, Na Zhang1
1Department of Urology, Tangshan Central Hospital, East of Guangming Road, South of Longfu South Road, West of Youyi Road West Auxiliary Road, North of Changning Road, Tangshan, 063000, China.
Objective:
Our study aims to investigate the mechanism of the miR-129-5p/SPN axis in clear cell renal cell carcinoma (ccRCC), providing a novel direction for the targeted therapy of ccRCC.
Methods:
Bioinformatics methods were implemented to find the differentially expressed genes (DEGs) associated with ccRCC from TCGA database. qRT-PCR was performed to detect miR-129-5p and SPN mRNA expression, while western bot was carried out for the detection of protein expression of SPN. Bioinformatics analysis was used to predict the binding sites of miR-129-5p on SPN 3'UTR, while dual-luciferase assay was conducted to verify their binding relationship. CCK-8 assay, colony formation assay, wound healing assay and Transwell assay were employed to measure ccRCC cell proliferative ability, cell formation ability, cell migratory and invasive abilities. Flow cytometry was implemented to assess cell cycle and apoptosis.
Results:
miR-129-5p exhibited a significantly down-regulated expression level in ccRCC, while SPN showed a remarkably up-regulated expression level. Overexpressed miR-129-5p inhibited ccRCC cell proliferative, invasive and migratory capacities while induced cell cycle arrest in G0/G1 phase and promoted cell apoptosis. Dual-luciferase assay confirmed that there was a binding relationship between miR-129-5p and SPN. Moreover, overexpressed miR-129-5p remarkably reduced SPN expression in cancer cells, weakened the promoting effect of SPN on cell proliferation, migration, invasion and cell cycle progress, and led to enhanced cell apoptotic activity.
Conclusions:
Our study proves the regulatory effect of the miR-129-5p/SPN axis in ccRCC, and provides a novel potential target for precise treatment of patients with ccRCC.
Insights
MicroRNA-129-5p (miR-129-5p) is downregulated in clear cell renal cell carcinoma (ccRCC), while SPN is upregulated. Restoring miR-129-5p inhibits ccRCC progression by targeting SPN, offering a new therapeutic strategy.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Clear cell renal cell carcinoma (ccRCC) is a prevalent kidney cancer subtype.
- Understanding the molecular mechanisms driving ccRCC progression is crucial for developing effective treatments.
Purpose of the Study:
- To investigate the role of the miR-129-5p/SPN axis in ccRCC.
- To explore the potential of targeting this axis for ccRCC therapy.
Main Methods:
- Bioinformatic analysis of TCGA data for differentially expressed genes.
- qRT-PCR and Western blot for gene and protein expression analysis.
- Dual-luciferase assay to confirm miR-129-5p and SPN interaction.
- Cell proliferation, migration, invasion, cell cycle, and apoptosis assays.
Main Results:
- miR-129-5p was significantly downregulated, and SPN was upregulated in ccRCC.
- Overexpression of miR-129-5p suppressed ccRCC cell proliferation, migration, and invasion.
- miR-129-5p directly targeted SPN, reducing its expression and downstream effects.
- miR-129-5p induced cell cycle arrest and promoted apoptosis in ccRCC cells.
Conclusions:
- The miR-129-5p/SPN axis plays a critical regulatory role in ccRCC.
- Targeting the miR-129-5p/SPN axis presents a promising therapeutic strategy for ccRCC patients.
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