Department of Laboratory, People's Hospital of Yuxi City, Yuxi 653100, P.R. China.
Department of Quality Control, Central Blood Station of Yuxi City, Yuxi 653100, P.R. China.
Biosci Rep. 2018 Aug 31;38(4). doi: 10.1042/BSR20171615.
Osteosarcoma (OS) is the most common histological form of primary bone cancer. It is most prevalent in teenagers and young adults. The present study aims at exploring the regulatory effect of microRNA-340 (miR-340) on OS cell proliferation, invasion, migration, and apoptosis via regulating the Notch signaling pathway by targeting β-catenin (cadherin-associated protein) 1 (CTNNB1). OS tissues belonging to 45 patients and normal femoral head tissues of 45 amputees were selected. Cells were allocated to different groups. hybridization was performed to determine the positive rate of miR-340 expression while immunohistochemistry was used to determine that of CTNNB1 and B-cell lymphoma 2 (Bcl-2). We used a series of experiments to measure the expressions of related factors and assess rates of cell proliferation, migration, invasion, cycle, and apoptosis respectively. Our results show that miR-340 was expressed a higher level in normal tissue than OS tissue. Expression of Notch, CTNNB1, hairy and enhancer of split 1 (Hes1), Bcl-2, Runt-related transcription factor 2 (Runx2), and osteocalcin increased and that of miR-340, Bcl-2 interacting mediator of cell death (BIM), and Bcl-2 associated protein X (Bax) decreased in OS tissues. U-2OS cell line had the highest miR-340 expression. We also found that the up-regulation of miR-340 had increased expression of miR-340, BIM, and Bax but decreased expression of Notch, CTNNB1, Hes1, Bcl-2, Runx2, and osteocalcin. Up-regulation of miR-340p lead to increased cell apoptosis, suppressed cell proliferation, migration, and invasion. Our study demonstrates that overexpression of miR-340 could suppress OS cell proliferation, migration, and invasion as well as promoting OS cell apoptosis by inactivating the Notch signaling pathway via down-regulating CTNNB1. Functional miR-340 overexpression might be a future therapeutic strategy for OS.
骨肉瘤（OS）是原发性骨癌最常见的组织学形式。它最常见于青少年和年轻人。本研究旨在通过靶向β-连环蛋白（钙粘蛋白相关蛋白）1（CTNNB1）来探索 microRNA-340（miR-340）对 OS 细胞增殖、侵袭、迁移和凋亡的调节作用，从而调节 Notch 信号通路。选择了 45 名患者的 OS 组织和 45 名截肢者的正常股骨头组织。将细胞分配到不同的组中。进行杂交以确定 miR-340 表达的阳性率，同时进行免疫组织化学以确定 CTNNB1 和 B 细胞淋巴瘤 2（Bcl-2）的表达。我们使用一系列实验分别测量相关因素的表达并评估细胞增殖、迁移、侵袭、周期和凋亡的比率。我们的结果表明，miR-340 在正常组织中的表达水平高于 OS 组织。Notch、CTNNB1、Hairy 和 Enhancer of Split 1（Hes1）、Bcl-2、Runt-related transcription factor 2（Runx2）和骨钙素的表达增加，而 miR-340、Bcl-2 相互作用的细胞死亡介体（BIM）和 Bcl-2 相关蛋白 X（Bax）的表达在 OS 组织中降低。U-2OS 细胞系具有最高的 miR-340 表达。我们还发现，miR-340 的上调增加了 miR-340、BIM 和 Bax 的表达，而降低了 Notch、CTNNB1、Hes1、Bcl-2、Runx2 和骨钙素的表达。miR-340p 的上调导致细胞凋亡增加，抑制细胞增殖、迁移和侵袭。我们的研究表明，过表达 miR-340 通过下调 CTNNB1 使 Notch 信号通路失活，从而抑制 OS 细胞增殖、迁移和侵袭，并促进 OS 细胞凋亡。功能性 miR-340 过表达可能是 OS 的一种未来治疗策略。
