Biological function of microRNA-30c/SOX9 in pediatric osteosarcoma cell growth and metastasis

X.-D. Zhang, Y.-N. Wang, X.-Y. Feng, J.-Y. Yang, Y.-Y. Ge, W.-Q. Kong

Department of Orthopedics, Xuzhou Central Hospital, Xuzhou, China. wqingkong@163.com

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• Oncology

OBJECTIVE: Osteosarcoma is one of the commonest malignant bone tumors, which frequently occurs in children all over the world. To find out methods to improve the therapeutic effect of osteosarcoma, it is necessary to detect the functioning mechanism of miR-30c to regulate the proliferation and metastasis of osteosarcoma cell.

PATIENTS AND METHODS: In order to reveal the expression level of miR-30c, quantitative Real-time PCR (qRT-PCR) method was chosen. To evaluate cell viability and proliferation rates, colony formation and cell counting kit-8 (CCK8) assay were introduced. Based on cell migration and invasion assay, metastasis capacity of breast cancer cells was studied. Protein levels were measured by Western blotting assay and cell cycle distribution was identified by flow cytometry. Bioinformatics analysis and Luciferase assay were used to predict and verify the target gene.

RESULTS: Compared with pericarcinomatous tissues (n=38), miR-30c in osteosarcoma tissues was significantly suppressed. Overexpressed miR-30c could weaken osteosarcoma cell’s abilities of viability, proliferation, migration and invasion. Moreover, it could also encourage osteosarcoma cell apoptosis and block cell cycle at G0/G1 phase. According to bioinformatics analysis and Luciferase reporter assay, SOX9 was recognized as the target gene of miR-30c. Restoration of SOX9 could make miR-30c regain the ability of suppression on tumorigenesis of osteosarcoma cells.

CONCLUSIONS: MiR-30c could play an important role in tumor suppression for pediatric osteosarcoma development and metastasis by targeting SOX9 in vitro. Thus, a creative and potential target was provided for diagnosis and treatment of osteosarcoma.

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