[MiR-300 inhibits invasion and metastasis of osteosarcoma cell MG63 by negatively regulating PTTG1].

OBJECTIVE:To investigate the effects of miR-300 and on osteosarcoma invasion and metastasis and explore the molecular mechanism of osteosarcoma invasion and metastasis. OBJECTIVE:Western blot was used to detect the expression of duanyu1547G1 in human osteoblasts hFOB1.19 and osteosarcoma cell MG63 and to detect the transfection efficiency of cells transfected with plasmid; Transwell invasion assay and CCK8 assay detected the effects of knockdown of duanyu1547G1 and overexpression of miR-300 on the invasion and proliferation of osteosarcoma cell MG63. On-line prediction and screening of microRNAs (miRNAs) with complementary duanyu1547G1 binding was conducted. qRT-PCR was performed to examine the expression of miR-300 in hFOB1.19 and MG63 cells, and Western blotting was used to detect the expression of duanyu1547G1 in MG63 cells after transfection with a miR- 300 plasmid. Double luciferase assay was used to detect the targeted binding of miR-300 and Transwell invasion assay and CCK8 assay were used to detect the effects of overexpression of miR-300 and overexpression of duanyu1547G1 plasmid on invasion and proliferation of osteosarcoma cell line was highly expressed in MG63 cells (P=0.0002). duanyu1547G1 knockdown significantly inhibited the invasion (P=0.0002) and proliferation (P=0.0039) of MG63 cells. Based on the results of online prediction of complementary miRNAs to duanyu1547G1 and analysis of the data from NCBI database, miR-300 was determined as the target miRNA in this study. qRT-PCR results showed a significantly decreased expression of miR-300 in MG63 cells (P=0.0004). Overexpression of MiR-300 in MG63 cells significantly decreased the expression of duanyu1547G1 (P=0.0007), and the expressions of miR-300 and duanyu1547G1 were negatively correlated. Dual luciferase assay showed that miR-300 could specifically bind to duanyu1547G1 (P=0.001). Overexpression of duanyu1547G1 could significantly reverse the effect of miR-300 overexpression on invasion (P=0.0003) and proliferation (P=0.0077) of MG63 cells. OBJECTIVE:Overexpression of miR-300 can inhibit the invasion and metastasis of osteosarcoma cell MG63 by targeting

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