Department of Orthopedics, Southern Medical University, Guangzhou, Guangdong, China.
Eur Rev Med Pharmacol Sci. 2018 Mar;22(6):1602-1608. doi: 10.26355/eurrev_201803_14566.
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.
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.
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.
Our data provided the first evidence which supported the function of miR-384 as a tumor suppressor in OS by targeting SETD8.
有研究报道 miR-384 在多种癌症中下调表达，并发挥肿瘤抑制因子的作用。然而，miR-384 在骨肉瘤（OS）中的表达和功能尚未得到研究。在本研究中，我们旨在分析 miR-384 在 OS 进展中的作用和机制。
采用实时定量聚合酶链反应（qRT-PCR）检测 OS 组织和细胞中 miR-384 的表达。通过 MTT assay、集落形成分析和 Transwell 检测分析 miR-384 对人 OS 细胞的作用。采用 Western blot 分析 SETD8 的表达，应用荧光素酶报告基因检测分析 miR-384 在 OS 细胞中的靶基因。
我们发现 miR-384 在 OS 组织和 OS 细胞系中的表达明显低于相邻的非癌组织和正常骨细胞系。进一步的功能分析表明，miR-384 的上调显著抑制了 OS 细胞的增殖、迁移和侵袭，而 miR-384 的下调则对 OS 细胞具有相反的作用。此外，双荧光素酶报告基因检测、qRT-PCR 和 Western blot 鉴定 SETD8 为 miR-384 的潜在靶基因。最后，我们观察到上调 SETD8 逆转了过表达 miR-384 对 OS 细胞增殖、迁移和侵袭的影响。
我们的数据首次提供了证据，表明 miR-384 通过靶向 SETD8 在 OS 中作为肿瘤抑制因子发挥作用。
