miR-384 通过下调 SETD8 的表达抑制骨肉瘤细胞的生长和转移。
MicroRNA-384 downregulates SETD8 expression to suppress cell growth and metastasis in osteosarcoma cells.
机构信息
Department of Orthopedics, Southern Medical University, Guangzhou, Guangdong, China.
出版信息
Eur Rev Med Pharmacol Sci. 2018 Mar;22(6):1602-1608. doi: 10.26355/eurrev_201803_14566.
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.
目的
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。
患者与方法
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。
结果
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。
结论
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。
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