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微小RNA-451对肝脏受体同源物-1的抑制作用可抑制骨肉瘤细胞的增殖。

Suppression of liver receptor homolog-1 by microRNA-451 represses the proliferation of osteosarcoma cells.

作者信息

Li Zhiyong, Wu Shuwen, Lv Shouzheng, Wang Huili, Wang Yong, Guo Qiang

机构信息

Orthopaedic Department, Tianjin Baodi Hospital, Tianjin 301800, China.

Orthopaedic Department, Tianjin Baodi Hospital, Tianjin 301800, China.

出版信息

Biochem Biophys Res Commun. 2015 Jun 5;461(3):450-5. doi: 10.1016/j.bbrc.2015.04.013. Epub 2015 Apr 11.

Abstract

Liver receptor homolog-1 (LRH-1) plays an important role in the onset and progression of many cancer types. However, the role of LRH-1 in osteosarcoma has not been well investigated. In this study, the critical role of LRH-1 in osteosarcoma cells was described. Quantitative polymerase chain reaction and Western blot analysis results revealed that LRH-1 was highly overexpressed in osteosarcoma cells. LRH-1 was knocked down by small interfering RNA (siRNA), and this phenomenon significantly inhibited osteosarcoma cell proliferation. Bioinformatics analysis results showed that LRH-1 contained putative binding sites of microRNA-451 (miR-451); this result was further validated through a dual-luciferase activity reporter assay. miR-451 was overexpressed in osteosarcoma cells through transfection of miR-451 mimics; miR-451 overexpression then significantly inhibited LRH-1 expression and cell proliferation. The loss of LRH-1 by siRNA or miR-451 mimics significantly impaired Wnt/β-catenin activity, leading to G0/G1 cell cycle arrest. Results showed that LRH-1 is implicated in osteosarcoma. Therefore, miR-451-induced suppression of LRH-1 can be a novel therapy to treat osteosarcoma.

摘要

肝脏受体同源物-1(LRH-1)在多种癌症类型的发生和发展中起重要作用。然而,LRH-1在骨肉瘤中的作用尚未得到充分研究。在本研究中,描述了LRH-1在骨肉瘤细胞中的关键作用。定量聚合酶链反应和蛋白质印迹分析结果显示,LRH-1在骨肉瘤细胞中高度过表达。通过小干扰RNA(siRNA)敲低LRH-1,这种现象显著抑制了骨肉瘤细胞增殖。生物信息学分析结果表明,LRH-1含有微小RNA-451(miR-451)的假定结合位点;这一结果通过双荧光素酶活性报告基因检测进一步得到验证。通过转染miR-451模拟物在骨肉瘤细胞中过表达miR-451;miR-451过表达随后显著抑制LRH-1表达和细胞增殖。通过siRNA或miR-451模拟物使LRH-1缺失显著损害Wnt/β-连环蛋白活性,导致G0/G1细胞周期停滞。结果表明,LRH-1与骨肉瘤有关。因此,miR-451诱导的LRH-1抑制可能是一种治疗骨肉瘤的新疗法。

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