Department of Orthopedics, The Second Xiangya Hospital of Central South University, Changsha 410011, China.
Biosci Rep. 2020 Jun 26;40(6). doi: 10.1042/BSR20201232.
Osteosarcoma is a malignant bone tumour with the lowest survival rates out of all paediatric cancers and is primarily diagnosed in children and adolescents. MNAT1 is a subunit in the cyclin-dependent kinase-activating kinase complex. Abnormal up-regulation of MNAT1 has been associated with the poor prognosis of multiple cancers. Bioinformatics analysis showed that has-circ-0001146 and miR-26a-5p were involved in the regulation of MNAT1 in osteosarcoma. The present study investigated the regulatory effects of has-circ-0001146 and miR-26a-5p on MNAT1 expression using luciferase reporter and RNA-pull down assays. The effects of the has-circ-0001146/miR26a-5p/Mnat1 network on the proliferation and invasion of osteosarcoma were evaluated by cell viability, apoptosis, migration, and invasion assays. Osteosarcoma tissues showed higher MNAT1 and has-circ-0001146 expression than adjacent normal tissues, although the expression of MNAT1 was not significantly up-regulated in sarcomas according to TCGA databases. As indicated by luciferase reporter and RNA-pull down assays, miR-26a-5p was able to bind to both has-circ-0001146 and MNAT1 mRNA. The depletion of has-circ-0001146 as well as the increase of miR-26a-5p decreased MNAT1 expression in osteosarcoma cells, while the reduction of miR-26a-5p was associated with increased MNAT1 expression. These data suggested that has-circ-0001146 promoted MNAT1 expression by competitively binding to miR-26a-5p with MNAT1 mRNA. The depletion of has-circ-0001146 or MNAT1 or the increase of miR-26a-5p inhibited osteosarcoma cell viability and invasion, and increased apoptosis. Reduction of miR-26a-5p conversely promoted osteosarcoma cell viability and invasion. The present study confirmed that has-circ-0001146 blocked miR-26a-5p targeting MNAT1 in osteosarcoma cells, thereby promoting the malignant behaviours of osteosarcoma cells.
骨肉瘤是一种恶性骨肿瘤,是儿童癌症中存活率最低的肿瘤,主要发生于儿童和青少年。MNAT1 是细胞周期蛋白依赖性激酶激活激酶复合物的一个亚基。MNAT1 的异常上调与多种癌症的不良预后有关。生物信息学分析表明,has-circ-0001146 和 miR-26a-5p 参与了骨肉瘤中 MNAT1 的调节。本研究通过荧光素酶报告和 RNA 下拉测定,研究了 has-circ-0001146 和 miR-26a-5p 对 MNAT1 表达的调节作用。通过细胞活力、凋亡、迁移和侵袭测定,评估了 has-circ-0001146/miR26a-5p/Mnat1 网络对骨肉瘤增殖和侵袭的影响。骨肉瘤组织中 MNAT1 和 has-circ-0001146 的表达均高于相邻正常组织,尽管根据 TCGA 数据库,MNAT1 在肉瘤中的表达并没有明显上调。荧光素酶报告和 RNA 下拉测定表明,miR-26a-5p 能够与 has-circ-0001146 和 MNAT1 mRNA 结合。has-circ-0001146 的耗竭以及 miR-26a-5p 的增加均降低了骨肉瘤细胞中的 MNAT1 表达,而 miR-26a-5p 的减少则与 MNAT1 表达的增加有关。这些数据表明,has-circ-0001146 通过与 MNAT1 mRNA 竞争性结合 miR-26a-5p 促进 MNAT1 的表达。has-circ-0001146 或 MNAT1 的耗竭或 miR-26a-5p 的增加均抑制骨肉瘤细胞活力和侵袭,并增加凋亡。miR-26a-5p 的减少则相反地促进了骨肉瘤细胞的活力和侵袭。本研究证实,has-circ-0001146 阻断了骨肉瘤细胞中 miR-26a-5p 对 MNAT1 的靶向作用,从而促进了骨肉瘤细胞的恶性行为。