Han X G, Wang D W, Bi Z G, Gao F
Department of Orthopedics, First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, China.
Pharmacy Intravenous Admixture Service, First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, China.
J Biol Regul Homeost Agents. 2016 Apr-Jun;30(2):381-7.
This study was designed to investigate the regulatory effect of estrogen receptor-α (ERα)-mediated Wnt/β-catenin signaling pathway on osteoblast proliferation. Mc3T3-El cells were infected by ERα and ERβ small interfering ribose nucleic acid (siRNA) viruses and treated with estradiol 2 (E2) for 120 min after 24-h infection. Western blot was used to detect expressions of β-catenin, Gsk 3β, p-Gsk3β (Ser9) and CyclinDl; and methyl thiazolyl tetrazolium (MTT) was applied to detect osteoblast proliferation after interference by different ERs. Western blot results indicated that the expressions of β-catenin, p-Gsk3β (Ser9) and CyclinDl decreased after ERβ interference and ERα + ERβ joint interference, and a more obvious decrease was found after the joint interference. After ERβ interference, β-catenin, p-Gsk3β (Ser9) and CyclinDl were strongly expressed compared with expressions in the blank control group. MTT results demonstrated that the proliferation rate of osteoblast was lower after the joint interference than after ERβ interference, while a slight increase was found in the proliferation rate after ERβ interference in comparison with the blank control group. It can be concluded that estradiol is able to promote the proliferation of osteoblasts in mice by ERα-mediated Wnt/β-catenin signaling pathway.
本研究旨在探讨雌激素受体-α（ERα）介导的Wnt/β-连环蛋白信号通路对成骨细胞增殖的调控作用。用ERα和ERβ小干扰核糖核酸（siRNA）病毒感染Mc3T3-E1细胞，感染24小时后用雌二醇2（E2）处理120分钟。采用蛋白质免疫印迹法检测β-连环蛋白、糖原合成酶激酶3β（Gsk 3β）、磷酸化糖原合成酶激酶3β（p-Gsk3β，Ser9位点）和细胞周期蛋白D1（CyclinDl）的表达；采用甲基噻唑基四氮唑（MTT）法检测不同雌激素受体干扰后成骨细胞的增殖情况。蛋白质免疫印迹结果显示，干扰ERβ以及联合干扰ERα+ERβ后，β-连环蛋白、p-Gsk3β（Ser9位点）和CyclinDl的表达均降低，联合干扰后的降低更为明显。与空白对照组相比，干扰ERβ后β-连环蛋白、p-Gsk3β（Ser9位点）和CyclinDl的表达增强。MTT结果表明，联合干扰后成骨细胞的增殖率低于干扰ERβ后，而干扰ERβ后的增殖率与空白对照组相比略有升高。由此得出结论，雌二醇可通过ERα介导的Wnt/β-连环蛋白信号通路促进小鼠成骨细胞的增殖。
