Abstract
MicroRNA-206 has proven to be down-regulated in many human malignancies in correlation with tumour progression. Our study aimed to characterize miR-206 contributions to initiation and malignant progression of human osteosarcoma. MiR-206 expression was detected in human osteosarcoma cell line MG63, human normal osteoblastic cell line hFOB 1.19, and paired osteosarcoma and normal adjacent tissues from 65 patients using quantitative RT-PCR. Relationships of miR-206 levels to clinicopathological characteristics were also investigated. Moreover, miR-206 mimics and negative control siRNA were transfected into MG63 cells to observe effects on cell viability, apoptosis, invasion and migration. We found that miR-206 was down-regulated in the osteosarcoma cell line MG63 and primary tumor samples, and decreased miR-206 expression was significantly associated with advanced clinical stage, T classification, metastasis and poor histological differentiation. Additionally, transfection of miR-206 mimics could reduce MG- 63 cell viability, promote cell apoptosis, and inhibit cell invasion and migration. These findings indicate that miR-206 may have a key role in osteosarcoma pathogenesis and development. It could serve as a useful biomarker for prediction of osteosarcoma progression, and provide a potential target for gene therapy.
Highlights
Osteosarcoma (OS) is the most common human primary malignant bone tumor in children and young adults, which accounts for approximately 60% of malignant bone tumors in the first 2 decades of life (Geller and Gorlick, 2010)
We found that miR-206 was down-regulated in the osteosarcoma cell line MG63 and primary tumor samples, and decreased miR-206 expression was significantly associated with advanced clinical stage, T classification, metastasis and poor histological differentiation
Scratch migration assay When MG-63 cells transfected with negative control (NC) or miR-206 mimics were seeded and grown to confluence, a scratch was set with a pipette tip running though the dish and cultured under standard conditions for 24 h
Summary
Osteosarcoma (OS) is the most common human primary malignant bone tumor in children and young adults, which accounts for approximately 60% of malignant bone tumors in the first 2 decades of life (Geller and Gorlick, 2010). Subsequent reports have shown that miRNAs are differentially expressed in many cancers (Calin et al, 2004; Song et al, 2010). Recent studies have analyzed various miRNAs that might contributing to invasion and metastasis in OS (Kobayashi et al, 2012). Using high-throughput technology, such as miRNA oligonucleotide arrays and quantitative RT-PCR for validation, previous studies have corroborated aberrant miR-206 expression in human malignancies such as lung cancer (Wang et al, 2011), rhabdomyosarcoma (Missiaglia et al, 2010), breast cancer (Iorio et al, 2005), and endometrioid adenocarcinoma (Chen et al, 2012). In this study, the expression of miR-206 in OS tissue and cell line MG63 was examined by real-time PCR. The effects of miR-206 on MG63 cell proliferation, apoptosis, invasion and migration were investigated
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