基础医学与临床 ›› 2012, Vol. 32 ›› Issue (7): 761-766.

• 研究论文 • 上一篇    下一篇

Txndc5介导雌激素诱导的小鼠前成骨细胞MC3T3-E1增殖加快

王乐,卢雅彬,熊霞辉,朱宁,陈梅红   

  1. 中国医学科学院基础医学研究所
  • 收稿日期:2012-04-11 修回日期:2012-05-24 出版日期:2012-07-05 发布日期:2012-06-20
  • 通讯作者: 陈梅红 E-mail:chenmhxc@gmail.com
  • 基金资助:
    骨髓间充质干细胞瘤化中STAT3信号通路过度激活的作用机制与规避研究;骨髓间充质干细胞瘤化中STAT3信号通路过度激活的作用机制与规避研究

Txndc5 mediates estrogen accelerated proliferation of murine pre-osteoblast MC3T3-E1

  • Received:2012-04-11 Revised:2012-05-24 Online:2012-07-05 Published:2012-06-20
  • Contact: Mei-Hong CHEN E-mail:chenmhxc@gmail.com

摘要: 目的 研究Txndc5基因在小鼠前成骨细胞增殖中的作用。 方法 用雌激素诱导小鼠前成骨细胞系MC3T3-E1,或在MC3T3-E1细胞中分别用质粒载体过表达Txndc5和用siRNA抑制Txndc5的表达后,Western blot检测Txndc5和细胞周期蛋白水平,荧光实时定量PCR检测Cyclin A的mRNA水平,MTS法和细胞计数法检测细胞增殖速度,流式细胞术检测细胞周期。 结果 雌激素诱导MC3T3-E1增殖加快时,Txndc5的蛋白水平亦上升。抑制Txndc5的表达阻止雌激素的促细胞增殖作用。过表达Txndc5使MC3T3-E1细胞增殖速度加快,S期细胞比例增加,同步化细胞进入细胞周期18h时,过表达Txndc5组Cyclin A 的表达升高,且S期细胞比例为18.69%±4.08%,而对照组仅为8.15%±3.68%。抑制Txndc5的表达则使MC3T3-E1细胞增殖速度减慢,S期细胞比例减少,Cyclin A 的表达下降。抑制Cyclin A的表达减弱Txndc5的促细胞增殖作用。结论 Txndc5通过上调Cyclin A 的表达介导雌激素的促前成骨细胞增殖作用。

关键词: Txndc5, MC3T3-E1细胞, 细胞增殖, 雌激素

Abstract: Objective To investigate the role of Txndc5 in murine pre-osteoblast proliferation. Methods Murine pre-osteoblast cell line MC3T3-E1 was treated with estrogen. Txndc5 was either over-expressed by plasmid vector or knocked down by siRNA in MC3T3-E1 cell. The protein level of Txndc5 and Cyclins was evaluated by Western blot. The mRNA level of Cyclin A was measured by quantitative real-time PCR. Cell proliferation rate was determined by MTS assay and cell counting. Cell cycle distribution was revealed by flow cytometry analysis. Results The protein level of Txndc5 was up-regulated in estrogen-induced MC3T3-E1 proliferation. Knockdown of Txndc5 blocked estrogen-induced cell proliferation. Over-expression of Txndc5 accelerated MC3T3-E1 cell proliferation, increased the proportion of cells in S phase. Eighteen hours after the synchronized cells entered cell cycle, the expression of Cyclin A was up-regulated, and the percentage of cells in S phase was 18.69%±4.08% in cells over-expressing Txndc5, which was only 8.15%±3.68% in control cells. On the other hand, knockdown of Txndc5 inhibited the growth of MC3T3-E1 cells, decreased the proportion of cells in S phase, and down-regulated the expression of Cyclin A. Knockdown of Cyclin A attenuated the cell proliferation-enhancing effect of Txndc5. Conclusion Txndc5 mediates estrogen-accelerated pre-osteoblast proliferation through up-regulation of Cyclin A.

Key words: Txndc5, MC3T3-E1 cell, cell proliferation, estrogen

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