转录因子Twist 对小鼠成肌细胞C2C12 分化的影响

The influence of transcriptional factors Twist to mouse myoblast C2C12 differentiation process

转录因子Twist 是高度保守的碱性螺旋-环-螺旋蛋白家族B类成员,包括Twist 1 和Twist 2. Twist 在生长发育以及肿瘤的发生发展等过程都具有重要作用,但是其对小鼠成肌细胞系C2C12 分化过程的影响尚不明确.因此本研究探讨Twist 在C2C12分化过程中的表达情况及其对分化过程的影响.通过低血清诱导体外肌肉细胞分化模型,观察分化过程中细胞形态变化.采用Western blot 检测肌肉细胞分化标志分子myogenin 和MY-32 在C2C12细胞分化第1天, 2天, 4天的表达.检测分化过程中Twist 1 和Twist 2 转录和蛋白表达水平.结果显示C2C12 细胞分化过程中细胞形态逐渐变长变粗融合形成多核肌管. Myogenin 随着分化进程表达量升高, MY-32 在分化第4天出现显著表达.而Twist 1 在C2C12 细胞分化第2天和第4天转录水平均提高(P<0.05),而Twist 2在细胞分化第4天转录水平均显著提高(P<0.05),但是Twist 2蛋白表达在分化过程中未见显著变化.该研究表明Twist 1和Twist 2基因在C2C12 细胞中转录水平均升高,但是蛋白表达水平未见显著差异.

The transcription factor Twist is a highly conserved class B member of the basic helix-loop-helix protein family,including Twist 1 and Twist 2.Twist plays an important role in the process of cell growth,development and tumor development,but its role in the difterentiation of mouse myoblast cell line C2C12 is still unclear.Therefore,the studies on the expression and role of Twist in C2C12 differentiation were perfomied here.The C2C12 myoblasts were cultured in low serum-containing DMEM to induce differentiation,and the cell morphology during differentiation was observed.Western blot was applied to detect the expression of muscle differentiation markers,inyogenin and M Y-32.Westem blot and RT-PCR were perfonned to detect the transcription and protein expression of Twist 1 and T wist 2 during differentiation.The results showed that C2C12 cells became longer and thicker and then fused to fbnn multinucleated myotubes under stimulating.The expression of myogenin increases,and MY-32 appears until the 4^th day of differentiation.During differentiation,the transcription of Twist 1 elevated significantly in day 2^th and day 4^th differentiation(P<0.05).The transcription of Twist 2 elevated significantly in day 4th differentiation(P<0.05),but no significant Twist 2 protein changes.This study showed that both Twist 1 and Twist 2 transcription were increased,but based on western blot,there is no significant difference for Twist 1 and Twist 2 protein during differentiation of C2C12 cells.