Tmed2促进体外小鼠前成骨细胞MC3T3-E1增殖  

Tmed2 accelerates murine MC3T3-E1 pre-osteoblast proliferation in vitro

作　　者：熊元[1] 熊霞辉[1] 卢雅彬[1] 朱宁[1] 陈梅红[1] 

机构地区：[1]中国医学科学院基础医学研究所 北京协和医学院 基础医学院 生物化学与分子生物学系 医学分子生物学国家重点实验室,北京100005

出　　处：《基础医学与临床》2012年第7期751-755,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(90608023);中国医学科学院基础医学研究所医学分子生物学国家重点实验室自主研究课题(2060204)

摘　　要：目的研究Tmed2基因对小鼠前成骨细胞增殖的影响。方法 1)分别在小鼠MC3T3-E1细胞中过表达和抑制Tmed2,检测细胞增殖情况。2)用雌激素处理细胞后,检测细胞的增殖及Tmed2基因的表达量。荧光实时定量PCR检测mRNA水平,MTS法检测细胞活力和增殖,流式细胞术检测细胞周期,Western blot法检测蛋白水平。结果过表达Tmed2使MC3T3-E1细胞的增殖速度加快,细胞周期中S期细胞比例明显增加,且Cyclin A的表达升高。而抑制Tmed2基因表达使MC3T3-E1细胞的增殖速度减慢。雌激素处理使细胞增殖速度加快的同时,Tmed2基因的表达显著增高。结论 Tmed2通过上调Cyclin A的表达,使S期细胞比例增加,加快小鼠前成骨细胞MC3T3-E1的增殖。此外,Tmed2的表达受雌激素的调控,可能参与雌激素促进MC3T3-E1细胞增殖的作用。Objective To study the role of Tmed2 in murine pre-osteoblast proliferation. Methods 1 ）Over-ex- press and knock down Treed2, and the proliferation rate of MC3T3-E1 cell was then detected. 2） MC3T3-E1 cells were treated with β-Estradiol, cell number was then counted and the mRNA level of Treed2 was analyzed. Quanti- tative real-time PCR was used to measure the mRNA level, MTS assay was used to assess the cell proliferation rate, flow cytometry was used to figure out the cell cycle distribution, and the protein level was evaluated by Western blot. Results Over-expression of Treed2 accelerated MC3T3-E1 cell proliferation, and the proportion of cells in S phase markedly increased. The expression of Cyclin A also increased. On the contrary, Knockdown of Treed2 de- celerated MC3T3-E1 cell proliferation. In [3-Estradiol promoted MC3T3-E1 proliferation, the mRNA level of Treed2 remarkably increased. Conclusions Tmed2 accelerates murine pre-osteoblast MC3T3-E1 proliferation through up- regulating Cyclin A expression and therefore increasing the proportion of cells in S phase. In addition, the expres- sion of Tmed2 is regulated by β-Estradiol, indicating that Tmed2 is potentially involved in the process of MC3T3- E1 proliferation promoted by β-Estradiol.

关 键 词：Tmed2 MC3T3-E1细胞 细胞增殖 雌激素 

分 类 号：R363[医药卫生—病理学]