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miR-200bc/429 通过靶向PMP22抑制骨肉瘤细胞增殖和侵袭。

miR-200bc/429 Inhibits Osteosarcoma Cell Proliferation and Invasion by Targeting PMP22.

作者信息

Li Xiaodong, Jiang Han, Xiao Lianping, Wang Shusen, Zheng Jinxin

机构信息

Department of Orthopedics, Tianjin third central hospital, Tianjin, China (mainland).

出版信息

Med Sci Monit. 2017 Feb 24;23:1001-1008. doi: 10.12659/msm.900084.

Abstract

BACKGROUND MicroRNAs (miRNAs) are small non-coding RNAs which play a crucial role in diverse biological processes and could contribute to cancer development and progression. MiR-200bc/429 have been found to be aberrantly expressed in osteosarcoma (OS). However, the features of miR-200bc/429 in the tumorigenesis and progress of OS remain poorly understood. MATERIAL AND METHODS The miR-200bc/429 expression was firstly identified in human OS clinical samples and cell lines by quantitative real-time PCR (qRT-PCR). After transfection with miR-200bc/429 mimics or negative control in U2OS or MG63 cells, cell proliferation was measured by CCK-8 assay. Following that, wound-healing assay and Transwell invasion assay were performed to evaluate cell migration and invasion, respectively. Finally, luciferase reporter assay and Western blot analysis were performed to determine if peripheral myelin protein-22 (PMP22) is a direct target of miR-200bc/429. RESULTS Results revealed that miR-200bc/429 were significantly depressed in human OS tissues and cell lines by qRT-PCR. Then, restoration of miR-200bc/429 significantly inhibited cell proliferation (P<0.05) and invasion (P<0.05) in vitro. Luciferase reporter assay and Western blot analysis revealed that miR-200bc/429 could directly target PMP22 3' untranslated region (UTR) and inhibit its expression in U2OS and MG63 cells. CONCLUSIONS These findings suggest that miR-200bc/429 inhibit OS cells proliferation and invasion by targeting PMP22, and function as a tumor suppressor and may be a patent molecular marker as well as a potential target for OS therapy.

摘要

背景

微小RNA(miRNA)是一类小的非编码RNA,在多种生物学过程中发挥关键作用,可能与癌症的发生和发展有关。已发现miR-200bc/429在骨肉瘤(OS)中异常表达。然而,miR-200bc/429在OS肿瘤发生和进展中的特征仍知之甚少。

材料与方法

首先通过定量实时PCR(qRT-PCR)在人OS临床样本和细胞系中鉴定miR-200bc/429的表达。在U2OS或MG63细胞中转染miR-200bc/429模拟物或阴性对照后,通过CCK-8法检测细胞增殖。随后,分别进行伤口愈合试验和Transwell侵袭试验以评估细胞迁移和侵袭。最后,进行荧光素酶报告基因试验和蛋白质免疫印迹分析,以确定外周髓鞘蛋白22(PMP22)是否为miR-200bc/429的直接靶点。

结果

qRT-PCR结果显示,miR-200bc/429在人OS组织和细胞系中显著下调。然后,miR-200bc/429的恢复显著抑制了体外细胞增殖(P<0.05)和侵袭(P<0.05)。荧光素酶报告基因试验和蛋白质免疫印迹分析表明,miR-200bc/429可直接靶向PMP22的3'非翻译区(UTR)并抑制其在U2OS和MG63细胞中的表达。

结论

这些发现表明,miR-200bc/429通过靶向PMP22抑制OS细胞增殖和侵袭,起到肿瘤抑制作用,可能是骨肉瘤的一个潜在分子标志物及治疗靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0bf9/5338570/fb62dadc1375/medscimonit-23-1001-g001.jpg

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