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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。
Objective: To study the expression and significance of DLK2 (delta drosophila homolog-like 2 or delta like 2 homologue) in embryonic cartilage development. Methods: The expression of DLK2 in ATDC5 cell line was induced by normal differentiation. The ATDC5-DLK2 lentivirus overexpression line was established to detect the effect of DLK2 gene on the proliferation and differentiation of ATDC5 cells. The expression of DLK2 in C57BL / 6 mice was detected by immunohistochemistry The temporal and spatial expression of the knee joint in mouse embryos (E11.5 d, E16.5 d, E18.5 d) and postnatal days (P7 d and P14 d) were analyzed using the SPSS16.0 software package for paired t-test. Results: The m RNA and protein levels of DLK2 decreased gradually during ATDC5 cell differentiation. The proliferation rate of ATDC5 cells was significantly increased after overexpression of DLK2 (P <0.05). However, ATDC5 cells were differentiated into cartilage markers (Col2a1, Acan, Col10a1 (P <0.05). At the early stage of embryonic development (E11.5 d), DLK2 began to express weakly at the cartilage formation site and gradually increased in the process of cartilage development, and DLK2 was not mature And mature chondrocytes. The expression of DLK2 on the surface of articular cartilage gradually decreased (P <0.05), but the depth was almost unchanged (P> 0.05). Conclusion: DLK2 may be involved in embryonic cartilage development of an important cytokine.