中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (10): 1808-1812.doi: 10.3969/j.issn.1673-8225.2012.10.022

• 干细胞转基因表达 transgenic expression in stem cells • 上一篇    下一篇

Dlx2基因过表达与前成骨细胞系MC3T3-E1成骨分化过程中的细胞凋亡和周期调控*****★◆

孙  昊，王旭东，代杰文，卢境婷，沈国芳   

  1. 上海交通大学医学院附属第九人民医院口腔颌面外科，上海市口腔医学重点实验室，上海市  200011
  • 收稿日期:2011-06-04 修回日期:2011-09-09 出版日期:2012-03-04 发布日期:2012-03-04
  • 通讯作者: 王旭东，副教授，上海交通大学医学院附属第九人民医院口腔颌面外科，上海市口腔医学重点实验室，上海市 200011 xudongwang70@hotmail.com
  • 作者简介:孙昊★，男，1984年生，山东省菏泽市人，汉族，2011年上海交通大学医学院毕业，硕士，医师，主要从事颅颌面畸形的基础和临床研究工作。 sunhaohappy@yahoo.com.cn
  • 基金资助:

    上海市重点学科建设项目(S30206)；上海市科学技术委员会(基础重点)项目(10JC1408700)；上海市自然基金(10ZR1418000)；上海市卫生局资助项目(2009077)；上海交通大学医工(理)交叉研究基金(YG2010MS55)

Effects of Dlx2 overexpression on cell cycle regulation and apoptosis of preosteoblast cells

Sun Hao, Wang Xu-dong, Dai Jie-wen, Lu Jing-ting, Shen Guo-fang   

  1. Shanghai Key Laboratory of Stomatology, Department of Oral and Maxillofacial Surgery, Ninth People’s Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai  200011, China
  • Received:2011-06-04 Revised:2011-09-09 Online:2012-03-04 Published:2012-03-04
  • Contact: author: Wang Xu-dong, Associate professor, Shanghai Key Laboratory of Stomatology, Department of Oral and Maxillofacial Surgery, Ninth People’s Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200011, China xudongwang70@hotmail.com
  • About author:Sun Hao★, Master, Physician, Shanghai Key Laboratory of Stomatology, Department of Oral and Maxillofacial Surgery, Ninth People’s Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200011, China sunhaohappy@yahoo.com.cn
  • Supported by:

    Shanghai Leading Academic Discipline Project, No. S30206*; Research Fund of Science and Technology Commission of Shanghai City, No. 10JC1408700*; the Natural Science Foundation of Shanghai, No. 10ZR1418000*; Research Fund of Shanghai Municipal Health Bureau, No. 2009077*; Combined Engineering and Medicine Project of Shanghai Jiao Tong University, No. YG2010MS55*

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摘要：

背景：Dlx2基因在颅神经嵴细胞迁徙进入第一鳃弓过程和颅颌面骨骼发育中起重要作用，但Dlx2基因对成骨细胞分化过程中细胞凋亡和细胞周期调控的影响尚未见报道。
目的：观察Dlx2基因过表达对前成骨细胞系MC3T3-E1成骨分化过程中细胞凋亡和细胞周期调控的影响。
方法：构建反转录病毒pMSCV-puro-Dlx2并转染矿化诱导液培养下的MC3T3-E1细胞，构建稳定过表达Dlx2基因的细胞系MC3T3-E1-Dlx2。RT-PCR和Western blot验证Dlx2基因过表达细胞系的建立。Annexin V/PI双染色后流式细胞分选检测细胞凋亡，PI/RNase双染色后流式细胞分选检测细胞周期变化。
结果与结论：实验成功构建稳定过表达Dlx2基因的细胞系MC3T3-E1-Dlx2。发现Dlx2基因过表达促进细胞凋亡(P < 0.05)，同时阻滞细胞周期于G1/G0期(P < 0.05)，减低细胞增殖性，促进细胞分化行使功能。
关键词：成骨细胞分化；Dlx2基因；稳定过表达；细胞凋亡；细胞周期
doi:10.3969/j.issn.1673-8225.2012.10.022

关键词: 成骨细胞分化, Dlx2基因, 稳定过表达, 细胞凋亡, 细胞周期

Abstract:

BACKGROUND: Dlx2 plays an important role in the regulation of cranial neural crest cells migration and craniofacial skeleton development. However, the effects of Dlx2 on apoptosis and cell cycle in osteoblast differentiation are not reported.
OBJECTIVE: To investigate the effects of Dlx2 overexpression on cell cycle regulation and apoptosis in MC3T3-E1 preosteoblast cells differentiation.
METHODS: Anti-retroviral pMSCV-puro-Dlx2 was transfected into MC3T3-E1 cells which were cultured in mineralization induced fluid. MC3T3-E1-Dlx2 cell lines stably overexpressing Dlx2 were constructed. Cell lines of Dlx2 over-expression were determined by RT-PCR and western blot. Apoptosis was detected by flow cytometry after Annexin V/PI and PI/RNase staining and cell cycle changes were detected by flow cytometry after PI/RNase staining.
RESULTS AND CONCLUSION: MC3T3-E1-Dlx2 cell lines was successfully constructed. Cell cycle was blocked in G1/G0 phase (P < 0.05) and apoptosis was up-regulated (P < 0.05) in Dlx2 overexpression cell model. Dlx2 overexpression reduces cell proliferation and promotes cell differentiation to exercise functions.

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