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  

Effects of Dlx2 overexpression on cell cycle regulation and apoptosis of preosteoblast cells MC3T3-E1 during osteogenic differentiation process

作　　者：孙昊[1] 王旭东[1] 代杰文[1] 卢境婷[1] 沈国芳[1] 

机构地区：[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

出　　处：《中国组织工程研究》2012年第10期1808-1812,共5页Chinese Journal of Tissue Engineering Research

基　　金：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)~~

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。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.

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 

分 类 号：R394.2[医药卫生—医学遗传学]