Medical Research Center of Shengjing Hospital, China Medical University, Shenyang, 110004, China.
Cell Biol Int. 2018 Aug;42(8):965-974. doi: 10.1002/cbin.10962. Epub 2018 Apr 11.
Previous studies of the role of 17β-estradiol (E2) in myoblast differentiation have produced conflicting data. Therefore, this work aimed to determine the role of E2 on myoblast differentiation and specific myofiber formation. Murine C2C12 myoblasts were cultured in proliferation medium or differentiation medium/10 nM E2. The role of E2 on specific myosin heavy chain (MyHC) or estrogen receptor (ER) expression was examined using real-time quantitative RT-PCR (RT-qPCR). Transcriptome studies of E2 on myoblast differentiation were accomplished by microarray analyses. The expression levels of candidate genes from microarrays and four and a half LIM domains 1 (Fhl1) were detected with RT-qPCR. E2 in differentiation medium significantly up-regulated MyHC I expression, but exerted the opposite effects on MyHC II a, MyHC II b, and MyHC II d. Both ER-α and ER-β were decreased in differentiated C2C12, and E2 partially restored ER-β expression. Sixty-two up-regulated and 116 down-regulated genes treated by E2 were identified, and RT-qPCR validation results showed seven cytoskeletal genes (Myh8, Cenpe, Jak3, Obscn, Ldb3, Mybpc2, Col4a3bp), three genes related to ion channels (Kcnq1, Lrrc26, P2rx3) and Fhl1 transcript 2 were associated with the effects of E2 on myoblast differentiation. These findings suggested E2 helped slow type MyH I fiber formation and impeded fast 2A, 2X/D, and 2B fiber formation.
先前关于 17β-雌二醇（E2）在成肌细胞分化中的作用的研究得出了相互矛盾的数据。因此，本研究旨在确定 E2 对成肌细胞分化和特定肌纤维形成的作用。将鼠 C2C12 成肌细胞在增殖培养基或分化培养基/10 nM E2 中培养。使用实时定量 RT-PCR（RT-qPCR）检查 E2 对特定肌球蛋白重链（MyHC）或雌激素受体（ER）表达的作用。通过微阵列分析研究 E2 对成肌细胞分化的转录组。使用 RT-qPCR 检测微阵列和四个半 LIM 结构域 1（Fhl1）中候选基因的表达水平。分化培养基中的 E2 显著上调 MyHC I 的表达，但对 MyHC II a、MyHC II b 和 MyHC II d 产生相反的影响。分化的 C2C12 中的 ER-α 和 ER-β 均降低，E2 部分恢复了 ER-β 的表达。鉴定出 62 个上调和 116 个下调基因，RT-qPCR 验证结果显示 7 个细胞骨架基因（Myh8、Cenpe、Jak3、Obscn、Ldb3、Mybpc2、Col4a3bp）、3 个与离子通道相关的基因（Kcnq1、Lrrc26、P2rx3）和 Fhl1 转录本 2 与 E2 对成肌细胞分化的作用有关。这些发现表明，E2 有助于减缓 I 型 MyH 纤维的形成，并阻碍快速 2A、2X/D 和 2B 纤维的形成。
