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An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
MicroRNA-27a functions as an oncogene in human osteosarcoma by targeting CCNG1
Tao Lin1, Quanping Ma2, Yuefeng Zhang3
1Department of Orthopedics, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong 250000, P.R. China.
Abstract:
Osteosarcoma is the most common type of malignant tumor arising from bone in children and adolescents. Accumulating evidences have shown the aberrant expression of numerous miRNAs is associated with the development and metastasis of osteosarcoma. The present study was conducted to investigate miR-27a expression in osteosarcoma tissues and cells. In the present study, quantitative RT-qPCR was used to measure the expression levels of miRNA and mRNA in osteosarcoma tissues and cells. Transwell assays were used to detect the effects of miR-27a on the invasive and migratory potential of cells. Luciferase reporter and western blot analysis were conducted to confirm cyclin G1 (CCNG1) as the target gene of miR-27a. The results showed that miR-27a was significantly upregulated in human osteosarcoma tissues and cell lines. The western blot analysis revealed that the overexpression of miR-27a suppressed CCNG1 protein expression. Luciferase reporter assays confirmed that CCNG1 is a direct target of miR-27a in osteosarcoma cells. The results suggest that miR-27a downregulates CCNG1 expression in osteosarcoma and acts as an oncogene directly targeting CCNG1. Thus, the miR-27a/CCNGI axis is a potential therapeutic target for human osteosarcoma.
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