Egr1对小鼠成肌细胞C2C12分化的影响  

The Effect of Egr1 on C2C12 Differentiation

作　　者：崔亚凤 严云勤[1] 李树峰[1] 佟慧丽[1] 

机构地区：[1]东北农业大学生命科学学院细胞与发育生物学实验室,哈尔滨150030

出　　处：《中国细胞生物学学报》2018年第1期25-32,共8页Chinese Journal of Cell Biology

基　　金：国家转基因专项"高产优质转基因肉牛新品种培育"(批准号:2014ZX08007-002);黑龙江省博士后科研启动金(批准号:LBH-Q14025)资助的课题~~

摘　　要：早期生长反应蛋白1(early growth response protein 1,Egr1)作为转录因子在细胞增殖、分化及凋亡等许多生物学过程中都扮演着重要角色,但是其对小鼠成肌细胞C2C12分化的影响尚不明确。该研究采用Western blot和免疫荧光技术检测Egr1在C2C12细胞分化过程中的表达规律及定位。利用CRISPR(clustered regularly interspaced short palindromic repeats)/Cas9技术分别激活和抑制Egr1的表达,进而探讨Egr1对C2C12细胞分化的影响。结果显示,随着C2C12细胞分化的进行,Egr1的表达量在分化的第5 d达到峰值,随后呈下降趋势。Egr1表达定位于C2C12的细胞核与细胞质中,随着分化的进行,其在细胞核和细胞质中的表达量均显著升高。分别激活或抑制Egr1后,C2C12细胞肌管融合率以及肌肉分化标志分子肌细胞生成蛋白(myogenin,MYOG)和肌球蛋白重链2(myosin heavy chain 2,MYH2)水平均显著增加或降低。该研究结果表明,Egr1能够促进体外小鼠成肌细胞C2C12的分化。Egr1（early growth response protein 1）, as a transcription factor, plays important roles in many biological processes, such as cell proliferation, differentiation, apoptosis and so on. However, its effect on C2 C12 cells differentiation remains unclear. Western blot and immunofluorescence assays were used to determine the protein level and localization of Egr1 in C2 C12 during its differentiation. To explore its effect on C2 C12 differentiation, Egr1 expression was activated and inhibited using the CRISPR/Cas9 technique. Egr1 protein level reached the peak on the 5 th day of differentiation and then gradually decreased during C2 C12 differentiation. Egr1 localized in the cytoplasm and nucleus of C2 C12 cells. Egr1 level gradually increased when C2 C12 induced to differentiation. During the C2 C12 differentiation, myotube fusion rate, myogenin（MYOG） and myosin heavy chain（MYH2） levels increased or decreased significantly when Egr1 were up-regulated or down-regulated, respectively. In summary, the results showed that Egr1 promotes C2 C12 differentiation in vitro.

关 键 词：早期生长反应蛋白1 C2C12 细胞分化 

分 类 号：Q25[生物学—细胞生物学]