J-F Zhang1, G-Y Zhang, X-M Hu, Z-P Luo, Y-Z Ma. 1. Department of Orthopedics, Southern Medical University, Guangzhou, Guangdong, China. zjf3614777@126.com.
Abstract
OBJECTIVE: MiR-384 was reported to be downregulated and functioned as a tumor suppressor in several cancers. However, the expression and function of miR-384 in osteosarcoma (OS) have not been investigated. In the present study, we aimed to analyze the effect and mechanism of miR-384 in the progression of OS. PATIENTS AND METHODS: Quantitative Real-time polymerase chain reaction (qRT-PCR) was used to determine the expression of miR-384 in OS tissues and cells. MTT assay, colony formation analysis, Transwell assays were performed to analyze the role of miR-384 in human OS cells. Western blotting was applied to analyze the expression of SETD8, and the luciferase reporter assay was used to assess the target gene of miR-384 in OS cells. RESULTS: We found that miR-384 was significantly lowly expressed in OS tissues and OS cell lines compared with the adjacent noncancerous tissues and normal bone cell lines, respectively. Further functional analysis indicated that up-regulation of miR-384 significantly inhibited OS cells proliferation, migration, and invasion, but down-regulation of miR-384 had the opposite effects on OS cells in vitro. Moreover, SETD8 was identified as the potential target of miR-384 using dual luciferase assay, qRT-PCR and Western blot. Finally, we observed that upregulation of SETD8 reversed the effects of overexpressing of miR-384 on the proliferation, migration, and invasion of OS. CONCLUSIONS: Our data provided the first evidence which supported the function of miR-384 as a tumor suppressor in OS by targeting SETD8.
OBJECTIVE:MiR-384 was reported to be downregulated and functioned as a tumor suppressor in several cancers. However, the expression and function of miR-384 in osteosarcoma (OS) have not been investigated. In the present study, we aimed to analyze the effect and mechanism of miR-384 in the progression of OS. PATIENTS AND METHODS: Quantitative Real-time polymerase chain reaction (qRT-PCR) was used to determine the expression of miR-384 in OS tissues and cells. MTT assay, colony formation analysis, Transwell assays were performed to analyze the role of miR-384 in human OS cells. Western blotting was applied to analyze the expression of SETD8, and the luciferase reporter assay was used to assess the target gene of miR-384 in OS cells. RESULTS: We found that miR-384 was significantly lowly expressed in OS tissues and OS cell lines compared with the adjacent noncancerous tissues and normal bone cell lines, respectively. Further functional analysis indicated that up-regulation of miR-384 significantly inhibited OS cells proliferation, migration, and invasion, but down-regulation of miR-384 had the opposite effects on OS cells in vitro. Moreover, SETD8 was identified as the potential target of miR-384 using dual luciferase assay, qRT-PCR and Western blot. Finally, we observed that upregulation of SETD8 reversed the effects of overexpressing of miR-384 on the proliferation, migration, and invasion of OS. CONCLUSIONS: Our data provided the first evidence which supported the function of miR-384 as a tumor suppressor in OS by targeting SETD8.
Authors: G M Viera; K B Salomao; G R de Sousa; M Baroni; L E A Delsin; J A Pezuk; M S Brassesco Journal: Clin Transl Oncol Date: 2019-04-04 Impact factor: 3.405