MiR-494-3p induced by compressive force inhibits cell proliferation in MC3T3-E1 cells
MiR-494-3p induced by compressive force inhibits cell proliferation in MC3T3-E1 cells
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DOI：
10.1016/j.jbiosc.2015.02.006
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发表时间：
2015-10-01
影响因子：
2.8
通讯作者：
Yoshimoto, Katsuhiko
中科院分区：
文献类型：
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作者：
Iwawaki, Yuki;Mizusawa, Noriko;Yoshimoto, Katsuhiko
机械刺激调节基本的细胞过程，如增殖、分化和形态发生。我们试图确定microRNA（miRNA）的表达在压缩处理过程中改变的MC 3 T3-E1，前成骨细胞系。微阵列分析后的逆转录定量聚合酶链反应显示，在MC 3 T3-E1细胞中，294 Pa压力下持续24 h可增加miR-494- 3 p、miR-146 a-5 p、miR-210- 3 p和miR-1247- 3 p的水平。在这些miRNA中，发现miR-494- 3 p抑制MC 3 T3-E1细胞的细胞增殖。此外，与对照细胞相比，经受压缩力的细胞显示出较慢的细胞生长。成纤维细胞生长因子受体2（FGFR 2）和Rho相关卷曲螺旋激酶1（ROCK 1）的mRNA水平，预测为miR-494- 3 p的靶点，在MC 3 T3-E1细胞中通过压缩力或miR-494- 3 p模拟物的过表达而降低。此外，使用荧光素酶报告基因测定法确定Fgfr 2和Rock 1的3 '-非翻译区中miR-494- 3 p的结合位点。总之，压力影响了MC 3 T3-E1细胞中包括miR-494- 3 p在内的几种miRNA的表达。压力可能通过上调miR-494- 3 p，随后抑制FGFR 2和ROCK 1基因来抑制成骨细胞的细胞增殖。(C)2015年，日本生物技术学会。All rights reserved.
Mechanical stimuli regulate fundamental cell processes such as proliferation, differentiation, and morphogenesis. We attempted to identify microRNA (miRNA) whose expression is changed during compressive treatment in MC3T3-E1, a pre-osteoblastic cell line. Microarray analysis followed by reverse transcription-quantitative polymerase chain reaction revealed that compressive force at 294 Pa for 24 h in MC3T3-E1 cells increased levels of miR-494-3p, miR-146a-5p, miR-210-3p, and miR-1247-3p. Among these miRNAs, miR-494-3p was found to inhibit cell proliferation in MC3T3-E1 cells. Furthermore, cells subjected to compressive force showed slower cell growth compared with control cells. Levels of mRNA for fibroblast growth factor receptor 2 (FGFR2) and Rho-associated coiled-coil kinase 1 (ROCK1), which were predicted to be targets of miR-494-3p, were decreased by compressive force or overexpression of miR-494-3p mimics in MC3T3-E1 cells. Furthermore, binding sites of miR-494-3p within 3'-untranslated regions of Fgfr2 and Rock1 were determined using luciferase reporter assay. In conclusion, compressive force affected expressions of several miRNAs including miR-494-3p in MC3T3-E1 cells. Compressive force might inhibit cell proliferation in osteoblasts by up-regulating miR-494-3p followed by FGFR2 and ROCK1 gene repressions. (C) 2015, The Society for Biotechnology, Japan. All rights reserved.