Department of Orthopedic Surgery, First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China.
Mol Med Rep. 2018 Mar;17(3):4777-4784. doi: 10.3892/mmr.2018.8374. Epub 2018 Jan 4.
Osteosarcoma (OS) is the most common primary malignancy of the bone in teenagers and accounts for 20‑35% of all malignant primary bone tumors. Increasing evidence shows that microRNAs (miRNAs) are abnormally expressed in several types of human cancer. miRNAs are necessary to maintain the malignant phenotype of cancer cells and can function as either tumor suppressors or oncogenes. The present study aimed to measure the expression of miRNA‑129‑5p (miR‑129‑5p) in OS, determine the effects of miR‑129‑5p on the malignant behaviors of OS cells, and elucidate the molecular mechanism underlying the oncogenesis and progression of OS. The expression levels of miR‑129‑5p in OS tissues and cell lines were measured using reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) analysis. SAOS‑2 and U2OS cells were then transfected with miR‑129‑5p mimics or miR‑negative control. The effects of miR‑129‑5p on the proliferation, migration and invasion of SAOS‑2 and U2OS cells in vitro were then evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, Transwell migration assay and invasion assays, respectively. In addition, bioinformatics analysis, a luciferase reporter assay, and RT‑qPCR and western blot analyses were used to examine whether Rho‑associated protein kinase 1 (ROCK1) was a direct target of miR‑129‑5p. The mRNA expression of ROCK1 in OS tissues was detected using RT‑qPCR analysis, and the biological roles of ROCK1 in OS cells were also evaluated. The results showed that miR‑129‑5p was significantly downregulated in the OS tissues and cell lines. The re‑expression of miR‑129‑5p suppressed the cell proliferation, migration and invasion of OS cells. In addition, ROCK1 was confirmed as a direct target of miR‑129‑5p. The mRNA expression of ROCK1 was high in OS tissues and inversely correlated with the expression of miR‑129‑5p. The downregulation of ROCK1 inhibited the proliferation, migration and invasion of OS cells. These findings suggested that miR‑129‑5p inhibited cell proliferation, migration and invasion in the development of OS via the negative regulation of ROCK1. The miR‑129‑5p/ROCK1 axis may serve as an efficient target in cancer therapy.
骨肉瘤（OS）是青少年中最常见的原发性骨恶性肿瘤，占所有原发性骨肿瘤的 20-35%。越来越多的证据表明，微小 RNA（miRNA）在几种类型的人类癌症中异常表达。miRNA 对于维持癌细胞的恶性表型是必需的，并且可以作为肿瘤抑制因子或癌基因发挥作用。本研究旨在测量骨肉瘤组织和细胞系中 miRNA-129-5p（miR-129-5p）的表达水平，确定 miR-129-5p 对骨肉瘤细胞恶性行为的影响，并阐明骨肉瘤发生和进展的分子机制。采用逆转录-定量聚合酶链反应（RT-qPCR）分析测量 miR-129-5p 的表达水平。然后用 miR-129-5p 模拟物或 miR-阴性对照转染 SAOS-2 和 U2OS 细胞。分别通过 3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐（MTT）比色法、Transwell 迁移实验和侵袭实验评估 miR-129-5p 对 SAOS-2 和 U2OS 细胞体外增殖、迁移和侵袭的影响。此外，通过生物信息学分析、荧光素酶报告基因测定以及 RT-qPCR 和 Western blot 分析，检测 Rho 相关蛋白激酶 1（ROCK1）是否为 miR-129-5p 的直接靶标。采用 RT-qPCR 分析检测骨肉瘤组织中 ROCK1 的 mRNA 表达，并评估 ROCK1 在骨肉瘤细胞中的生物学作用。结果表明，miR-129-5p 在骨肉瘤组织和细胞系中显著下调。miR-129-5p 的重新表达抑制了骨肉瘤细胞的增殖、迁移和侵袭。此外，ROCK1 被确认为 miR-129-5p 的直接靶标。ROCK1 的 mRNA 表达在骨肉瘤组织中较高，与 miR-129-5p 的表达呈负相关。ROCK1 的下调抑制了骨肉瘤细胞的增殖、迁移和侵袭。这些发现表明，miR-129-5p 通过负调控 ROCK1 抑制 OS 细胞的增殖、迁移和侵袭，从而抑制 OS 的发展。miR-129-5p/ROCK1 轴可能成为癌症治疗的有效靶点。
