Katase Naoki, Terada Kumiko, Suzuki Takahiro, Nishimatsu Shin-ichiro, Nohno Tsutomu
Department of Molecular and Developmental Biology, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama, 701-0192, Japan.
BMC Cell Biol. 2015 Apr 30;16:13. doi: 10.1186/s12860-015-0061-9.
Skeletal muscle differentiation is a multistep, complex pathway in which several important signaling molecules are involved. Recently, microRNAs (miRNAs), endogenous non-coding small RNAs that regulate mRNAs, have been proposed to be involved in skeletal muscle differentiation. In this study, we identified skeletal muscle differentiation-associated miRNAs by comparing miRNA expression profiles between C2C12 cells and Wnt4 over-expressing C2C12 cells (W4-08), which can spontaneously differentiate into myotubes.
We identified miR-206, miR-133a, and miR-133b as up-regulated miRNAs and miR-487b, miR-3963 and miR-6412 as down-regulated miRNAs in differentiating cells. We focused on the down-regulated miRNAs because their functions were largely unknown. Transfection of mimics of these miRNAs into C2C12 cells resulted in significantly reduced expression of myogenic differentiation markers, including troponin T and myosin heavy chain fast type and slow type, but did not affect the expression of the myogenic transcription factors, MyoD and myogenin.
These miRNAs were characterized as new myogenic differentiation-associated miRNAs which may delay late myogenic differentiation or maturation.
骨骼肌分化是一个多步骤的复杂过程,涉及多种重要的信号分子。最近,有研究提出微小RNA(miRNA),即调控mRNA的内源性非编码小RNA,参与骨骼肌分化。在本研究中,我们通过比较C2C12细胞和过表达Wnt4的C2C12细胞(W4-08)之间的miRNA表达谱,鉴定出与骨骼肌分化相关的miRNA,其中W4-08细胞可自发分化为肌管。
我们鉴定出在分化细胞中,miR-206、miR-133a和miR-133b为上调的miRNA,miR-487b、miR-3963和miR-6412为下调的miRNA。我们关注下调的miRNA,因为它们的功能大多未知。将这些miRNA的模拟物转染到C2C12细胞中,导致肌原性分化标志物的表达显著降低,包括肌钙蛋白T以及快肌型和慢肌型肌球蛋白重链,但不影响肌原性转录因子MyoD和肌细胞生成素的表达。
这些miRNA被鉴定为新的与肌原性分化相关的miRNA,它们可能会延迟晚期肌原性分化或成熟。