The Role Of KLF4 In The Differentiation Of C2C12 Myogenic Cells

[Objective] To explore the role of KLF4 in the differentiation of C2C12 myogenic cells and preliminarily explore the mechanism. [Method] C2C12 myogenic cells were stably transfected with KLF4 eukaryotic expression constructs or control vector. The cells were grown to confluence and then the culture medium was replaced with DMEM containing 5% heat-inactivated horse serum to induce differentiation. The formation of multinucleated myotubes and measured the concentration of creatine kinase M isozyme (CKM, a differentiation marker for muscle cells)were observed during differentiation. CDNA chips containing 14000 mouse cDNAs were employed to investigate the influence of KLF4 overexpression on the genes expression. The differentially expressed genes after KLF4 transfection were further analyzed using bioinformatics databases and softwares such as MEDLINE and BibioSphere pathway edition. [Results] KLF4 over-expression accelerated the formation of multinucleated myotubes and enhanced the concentration of creatine kinase M isozyme during differentiation.250 genes were down-regulated in KLF4-overexpressed C2C12 cells, of which 155 were known genes; Whereas 355 genes were up-regulated, of which 255 were known genes, including 24 cell cycle-relevent genes.These 24 cell cycle-relevent genes belong to five different signal transduction pathways,among which Cdknla and Trp53/p53 genes and the p53 signal transduction pathway are most important. [Conclusion] KLF4 could accelerate the differentiation of C2C12 myogenic cells induced by horse serum. This role may be related to the regulation of the downstream cell cycle-relevent genes by KLF4.