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Updated: Oct 9, 2025
miRNA Expression Analyses in Prostate Cancer Clinical Tissues
Published on: September 8, 2015
[Hsa_circ_0005221 promotes prostate cancer progression through the miR-339-5p/STAT5a pathway]
Sai-Sai Chen1, Yu-Wei Zhang2, Ya-Li Wang3
1Research Institute of Urology, Southeast University School of Medicine, Nanjing, Jiangsu 211189, China.
Insights
Hsa_circ_0005221 promotes prostate cancer (PCa) progression by regulating the miR-339-5p/STAT5a pathway. This circular RNA enhances PCa cell proliferation, invasion, and migration while suppressing apoptosis.
Area of Science:
- Molecular Biology
- Oncology
- Gene Regulation
Background:
- Prostate cancer (PCa) remains a significant health concern.
- Understanding the molecular mechanisms driving PCa progression is crucial for developing effective therapies.
- Circular RNAs (circRNAs) have emerged as key regulators in various cancers, including PCa.
Purpose of the Study:
- To elucidate the role of hsa_circ_0005221 in PCa progression.
- To investigate the molecular pathway involving hsa_circ_0005221, miR-339-5p, and STAT5a in PCa.
- To determine the therapeutic potential of targeting this pathway.
Main Methods:
- Quantitative real-time PCR (qPCR) to assess gene expression.
- Nuclear-cytoplasmic isolation to determine cellular localization.
- RNA pull-down and luciferase reporter assays to confirm molecular interactions.
- Cell transfection with plasmids and siRNAs to manipulate gene expression and assess functional changes (proliferation, invasion, migration, apoptosis, epithelial-mesenchymal transition).
Main Results:
- Hsa_circ_0005221 expression is significantly upregulated in PCa tissues and cells.
- Hsa_circ_0005221 and miR-339-5p are primarily located in the cytoplasm.
- Hsa_circ_0005221 promotes PCa cell proliferation, invasion, migration, and epithelial-mesenchymal transition (EMT) while inhibiting apoptosis.
- Hsa_circ_0005221 directly binds to miR-339-5p, and this interaction influences STAT5a expression.
- Knockdown of hsa_circ_0005221 or overexpression of miR-339-5p reversed the pro-cancerous effects.
Conclusions:
- Hsa_circ_0005221 acts as an oncogenic circRNA in PCa.
- The hsa_circ_0005221/miR-339-5p/STAT5a axis is a critical pathway driving PCa progression.
- Targeting hsa_circ_0005221 may represent a novel therapeutic strategy for PCa.
Objective:
To investigate the molecular mechanism of hsa_circ_0005221 regulating the progression of PCa through the miR-339-5p/STAT5a pathway.
Methods:
Localizations of hsa_circ_0005221 and miR-339-5p in cells were detected by nuclear-cytoplasmic isolation. MiRNA-339-5p was selected as the target miRNA bound to hsa_circ_0005221 by RNA pull-down assay. The binding site of the luciferase reporter gene was predicted by software and the binding capability of miR-339-5p validated by luciferase assay. The expression of hsa_circ_0005221 in the prostatic epithelial and PCa cells was determined by qPCR. The hsa_circ_0005221-overexpressed plasmid and siRNA were transfected into the PCa cells for measurement of their proliferation, invasion and migration abilities and the levels of epithelial-mesenchymal transformation (EMT) and apoptosis. After knockdown of hsa_circ_0005221 and transfection of miR-339-5p mimics and miR-339-5p inhibitor, the proliferation, invasion and migration abilities of the DU145 and LNCaP cells were detected, and so were the levels of the EMT signature protein, STAT5a and cell apoptosis.
Results:
The expression of hsa_circ_0005221 was significantly higher in the PCa than in the prostatic epithelial cells. Nuclear-cytoplasmic isolation experiments showed that hsa_circ_0005221 and miR-339-5p were mainly located in the cytoplasm. The proliferation, invasion and migration abilities and EMT were decreased and the apoptosis increased in the DU145 and LNCaP cells with knockdown of hsa_circ_0005221, which was just the reverse in those with overexpressed hsa_circ_0005221. Among the top 5 miRNAs predicted by software, miR-339-5p, miR-17 and miR-520h were shown by pull-down assay to be bound to hsa_circ_0005221, with most obvious changes in miR-339-5p when hsa_circ_0005221 knocked down or overexpressed. Luciferase reporter gene assay showed the binding of hsa_circ_0005221 to miR-339-5p. Knockdown of hsa_circ_0005221 and transfection of miR-339-5p mimics into the DU145 and LNCaP cells significantly reduced the proliferation, invasion and migration abilities of the cells and the N-cad level, increased their apoptosis and E-cad level, and up-regulated the expression of STAT5a, while overexpression of hsa_circ_0005221 and transfection of miR-339-5p mimics induced just the opposite effects.
Conclusions:
Hsa_circ_0005221 enhances the progression of prostate cancer through the miR-339-5p/STAT5a pathway.
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