DLK2基因在胚胎软骨发育中的表达及意义  

Expression and significance of DLK2 gene during cartilage development

作　　者：徐伟峰[1] 沈佩[1] 范宝婷 焦子先[1] 李慧萍[1] 张善勇[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院.口腔医学院口腔外科上海市口腔医学重点实验室,上海200011

出　　处：《中国口腔颌面外科杂志》2016年第4期302-307,共6页China Journal of Oral and Maxillofacial Surgery

基　　金：国家自然科学基金(81371168)

摘　　要：目的:研究DLK2(delta drosophila homolog-like 2或delta like 2 homologue)在胚胎软骨发育中的表达及意义。方法:检测ATDC5细胞系正常诱导分化过程中DLK2的表达改变;建立ATDC5-DLK2慢病毒过表达细胞系,检测DLK2基因对ATDC5细胞增殖及分化的影响;通过免疫组织化学检测DLK2在C57BL/6小鼠胚胎(E11.5 d,E16.5 d,E18.5 d)和出生后(P7 d和P14 d)膝关节的时空表达,采用SPSS16.0软件包对数据进行配对t检验。结果:ATDC5细胞正常诱导分化过程中,DLK2的m RNA和蛋白水平逐渐下降;过表达DLK2后,ATDC5细胞增殖速率显著提高(P<0.05),但ATDC5细胞成软骨标志物(Col2a1、Acan、Col10a1)的m RNA水平显著下降(P<0.05);胚胎发育早期(E11.5 d),DLK2在软骨形成处开始有较弱表达,在软骨发育过程中,其表达逐渐增加,而且DLK2在未成熟和成熟的软骨细胞均有表达,关节软骨表层的DLK2表达逐渐下降(P<0.05),但深层几乎不变(P>0.05)。结论:DLK2可能是参与胚胎软骨发育过程的一个重要细胞因子。PURPOSE： To study the expression and significance of DLK2 during embryonic cartilage development.METHODS： The expression of DLK2 was detected during the differentiation of ATDC5 cells. ATDC5-DLK2 cells which stably overexpressed DLK2 was constructed by lentivirus transfection, and the effect of DLK2 on the proliferation and differentiation of ATDC5 cells was studied. The temporal and spatial expression of DLK2 in cartilage of C57BL/6 mouse at embryos（E11.5,E16.5,E18.5 days） and postnatal time（P7 and P14 day） was examined by immunohistochemistry. The data was analyzed by paired t test using SPSS16.0 software package. RESULTS： The m RNA and protein levels of DLK2 decreased during the differentiation of ATDC5 cells; ATDC5-DLK2 cell line which stably ovexpressed DLK2 was established and identified, and DLK2 overexpression promoted the proliferation of ATDC5 cells（ P〈0.05）,but significantly inhibited the expression of chondrogenic marker genes（col2a1, acan, col10a1）（ P〈0.05）. DLK2 showed a weak expression in cartilage of early embryo（E11.5 days）, while gradually increased during the development of cartilage. DLK2 was expressed in both immature and mature chondrocytes, but the expression of DLK2 in articular cartilage surface decreased（P〈0.05）, while almost unchanged in the deep of articular cartilage（P〈0.05）. CONCLUSIONS： DLK2 may be an important factor involved in cartilage development.

关 键 词：DLK2 ATDC5细胞 软骨发育 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学]