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Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis
Published on: March 30, 2019
MicroRNA-132 inhibits cell growth and metastasis in osteosarcoma cell lines possibly by targeting Sox4
Yulong Liu1, Ye Li1, Jingchen Liu1
1Department of Orthopaedic Surgery, China-Japan Union Hospital of Jilin University, Changchun 130031, P.R. China.
Abstract:
Increasing evidence has confirmed that dysregulation of microRNAs (miRNAs) can contribute to the progression and metastasis of human tumors. Previous studies have shown that dysregulation of microRNAs (miRNAs) can contribute to the progression and metastasis of human tumors. However, the precise mechanisms of miR‑132 in osteosarcoma have not been well clarified. Real-time PCR was performed to detect the expression of miR‑132 in osteosarcoma cell lines. miR-132 mimic, miR‑132 inhibitor and negative control were transfected into osteosarcoma cells and the effects of miR‑132 on the cell growth and metastasis were investigated. Furthermore, protein level of Sox4 was measured by western blotting. Luciferase assays were performed to validate Sox4 as miR‑132 target in osteosarcoma cells. We found that miR‑132 was downregulated in osteosarcoma cell lines. Introduction of miR‑132 significantly inhibited proliferation, arrested cell cycle and induced apoptosis in osteosarcoma cells. Besides, invasion and epithelial-mesenchymal transition (EMT) of osteosarcoma cells was suppressed by overexpressing miR‑132. However, downregulation of miR‑132 promoted cell growth and metastasis in osteosarcoma cells. Bioinformatics analysis predicted that Sox4 was a potential target gene of miR‑132. Luciferase reporter assay demonstrated that miR‑132 could directly target Sox4. Moreover, the low level of miR‑132 was associated with increased expression of Sox4 in osteosarcoma cells. Sox4 inhibition suppressed cell malignant behaviors. Overexpression of Sox4 in osteosarcoma cells transfected with miR‑132 mimic partially reversed the inhibitory effect of miR‑132. In conclusion, miR‑132 inhibited cell growth and metastasis in osteosarcoma cells by downregulation of Sox4, and knockdown of Sox4 was essential for the miR-132-inhibited cell growth and metastasis in osteosarcoma cells.
Insights
MicroRNA-132 (miR-132) inhibits osteosarcoma growth and metastasis by targeting Sox4. Downregulation of miR-132 promotes tumor progression, while Sox4 knockdown is crucial for miR-132
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- MicroRNA (miRNA) dysregulation is linked to cancer progression and metastasis.
- The specific role of miR-132 in osteosarcoma pathogenesis remains unclear.
Purpose of the Study:
- To elucidate the precise mechanisms of miR-132 in osteosarcoma.
- To investigate the regulatory relationship between miR-132 and Sox4 in osteosarcoma cells.
Main Methods:
- Real-time PCR to quantify miR-132 expression.
- Transfection of miR-132 mimics/inhibitors and Sox4 siRNA.
- Western blotting for protein analysis.
- Luciferase assays to confirm direct targeting.
Main Results:
- miR-132 was significantly downregulated in osteosarcoma cell lines.
- Overexpression of miR-132 inhibited proliferation, induced apoptosis, and suppressed invasion and epithelial-mesenchymal transition (EMT).
- miR-132 directly targets Sox4, and low miR-132 levels correlate with high Sox4 expression.
Conclusions:
- miR-132 acts as a tumor suppressor in osteosarcoma by downregulating Sox4.
- Sox4 inhibition is essential for the anti-tumor effects of miR-132 in osteosarcoma.
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