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MicroRNA-18a 通过靶向 MED27 抑制骨肉瘤细胞生长并诱导细胞凋亡。

MicroRNA-18a inhibits cell growth and induces apoptosis in osteosarcoma by targeting MED27.

机构信息

Department of Pediatric Orthopaedics, Xinhua Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200092, P.R. China.

Department of Good Clinical Practice, Shanghai Tenth People's Hospital of Tong Ji University, Shanghai 200072, P.R. China.

出版信息

Int J Oncol. 2018 Jul;53(1):329-338. doi: 10.3892/ijo.2018.4374. Epub 2018 Apr 16.

Abstract

Osteosarcoma (OS) is a common malignant primary bone tumor and patients with OS are known to have a poor response to chemotherapy. MicroRNAs (miRNAs or miRs) are small non-coding RNA molecules (approximately 22 nucleotides in length) and they have recently become a topic for research as regards their role in cancer therapeutics. Previous studies have reported miR‑18a expression in patients with OS is significantly decreased compared with that in normal adjacent tissue. miR‑18a belongs to the miR‑17‑92 cluster encoded by the host gene MIR17HG. However, the detailed role of miR‑18a in OS remains to be determined. In this study, we demonstrated that miR‑18a mimics inhibited MG63 and Saos‑2 cell viability and migration. In addition, flow cytometry assay revealed that miR‑18a induced OS cell apoptosis. Western blot analysis indicated that the expression levels of Bcl‑2 and p‑Akt were downregulated, while the levels of cleaved caspase‑3 and Bax proteins were upregulated by miR‑18a. Moreover, we demonstrated that mediator complex subunit 27 (MED27) was the target of miR‑18a through dual luciferase assay. Finally, data from in vivo experiments indicated that tumor growth in mice was significantly suppressed by miR‑18a mimics, accompanied by a decrease in the percentage of Ki67-positive cells, and by the downregulation in MED27 and p‑Akt protein expression levels. The findings of the present study may aid in the clarification of the function of miR‑18a, particularly as regards its role in the regulation of OS cell apoptosis, and indicate that MED27 may be a potential novel therapeutic target in the treatment of OS.

摘要

骨肉瘤(OS)是一种常见的恶性原发性骨肿瘤,已知 OS 患者对化疗的反应较差。微小 RNA(miRNA 或 miR)是小的非编码 RNA 分子(约 22 个核苷酸长),最近它们在癌症治疗中的作用成为研究的主题。先前的研究报告称,骨肉瘤患者 miR-18a 的表达明显低于正常相邻组织。miR-18a 属于由宿主基因 MIR17HG 编码的 miR-17-92 簇。然而,miR-18a 在 OS 中的详细作用仍有待确定。在这项研究中,我们证明了 miR-18a 模拟物抑制了 MG63 和 Saos-2 细胞的活力和迁移。此外,流式细胞术分析显示 miR-18a 诱导 OS 细胞凋亡。Western blot 分析表明,miR-18a 下调了 Bcl-2 和 p-Akt 的表达水平,而上调了 cleaved caspase-3 和 Bax 蛋白的表达水平。此外,我们通过双荧光素酶报告实验证明 MED27 是 miR-18a 的靶基因。最后,体内实验数据表明,miR-18a 模拟物显著抑制了小鼠肿瘤的生长,伴随着 Ki67 阳性细胞比例的降低,以及 MED27 和 p-Akt 蛋白表达水平的下调。本研究的结果可能有助于阐明 miR-18a 的功能,特别是在调节 OS 细胞凋亡方面的作用,并表明 MED27 可能是治疗 OS 的潜在新治疗靶点。

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