17β-雌二醇调控大鼠成肌细胞L6 GNR4细胞迁移能力的机制研究

摘要

Objective To investigate the effect of 17β-estradiol on the migration of L6GNR4 myoblast and discuss the molecular mechanism. Methods L6GNR4 cells were cultured in high-glucose DMEM containing 10 nM 17β-estradiol and/or 50 nM tamoxifen. The effect of 17β-estradiol on the migration of L6GNR4 myoblasts was detec-ted by Wound-healing assays at 0 h,12 h and 24 h after medication. The expression levels of ER and key genes in-volved in myoblast migration (Pax3,Fhl1,Fhl1 interactor-Actg1 and Myh10) were examined by real-time RT-PCR in each group. Results 17β-estradiol inhibited the migration of myoblast after 24 h and down-regulated the expression of ERβ,Pax3,Fhl1 and Myh10 which could be totally or partly reversed by tamoxifen,while the same dose of estrogen has no obvious effect on Actg1 expression in L6GNR4. Conclusion 17β-estradiol can inhibit the expression of Pax3, Fhl1 and Myh10 by ERβ-mediated transcription,thus restraining the L6GNR4 myoblast migration.%目的：研究17β-雌二醇对大鼠成肌细胞 L6GNR4细胞迁移能力的调控作用及机制。方法L6GNR4细胞培养在含有10%胎牛血清的DMEM高糖增殖培养基中,培养基中添加10 nM雌激素17β雌二醇和/或50 nM雌激素竞争性拮抗剂Tamoxifen。应用细胞划痕试验在药物处理后0、12、24 h观察17β雌二醇对成肌细胞迁移能力的影响。在药物处理24 h后,应用荧光定量PCR技术检测各组细胞中雌激素受体α和β的表达。同时还检测了成肌细胞迁移相关关键基因(Pax3、Fhl1以及Fhl1相互作用蛋白Actg1、Myh10)在各组细胞中的表达情况。结果与对照组比较,17β-雌二醇在划痕24 h时可以明显抑制L6GNR4的细胞迁移。17β-雌二醇能够明显抑制ERβ的表达,而雌激素竞争性拮抗剂 Tamoxifen 能够部分逆转17β-雌二醇对 ERβ的抑制作用。17β-雌二醇能够明显抑制成肌细胞迁移相关的关键因子Pax3、Fhl1以及Fhl1相互作用蛋白Myh10的表达,而雌激素竞争性拮抗剂Tamoxifen能够逆转或部分逆转17β-雌二醇对Pax3、Fhl1和Myh10的转录抑制作用。相同浓度的雌激素对L6GNR4细胞中Actg1的表达改变不明显。结论雌激素17β-雌二醇通过ERβ转录抑制Pax3、Fhl1以及Fhl1的相互作用蛋白Myh10的表达,发挥其抑制大鼠成肌细胞迁移的作用。