title: Ten-Eleven Translocation-2 (Tet2) Is Involved in Myogenic Differentiation of Skeletal Myoblast Cells in Vitro.
abstract: Muscle cell differentiation is a complex process that is principally governed by related myogenic regulatory factors (MRFs). DNA methylation is considered to play an important role on the expression of MRF genes and on muscle cell differentiation. However, the roles of enzymes specifically in myogenesis are not fully understood. Here, we demonstrate that Tet2, a ten-eleven translocation (Tet) methylcytosine dioxygenase, exerts a role during skeletal myoblast differentiation. By using an immunostaining method, we found that the levels of 5-hydroxymethylcytosine (5-hmC) were much higher in differentiated myotubes than in undifferentiated C2C12 myoblasts. Both Tet1 and Tet2 expression were upregulated after differentiation induction of C2C12 myoblasts. Knockdown of Tet2, but not Tet1, significantly reduced the expression of myogenin as well as Myf6 and myomaker, and impaired myoblast differentiation. DNA demethylation of myogenin and myomaker promoters was negatively influenced by Tet2 knockdown as detected by bisulfite sequencing analysis. Furthermore, although vitamin C could promote genomic 5hmC generation, myogenic gene expression and myoblast differentiation, its effect was significantly attenuated by Tet2 knockdown. Taken together, these results indicate that Tet2 is involved in myoblast differentiation through promoting DNA demethylation and myogenic gene expression.
authors.affiliation: State Key Laboratory of Agrobiotechnology and College of Biological Science, China Agricultural University, Beijing, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Zhong
authors.affiliation: State Key Laboratory of Agrobiotechnology and College of Biological Science, China Agricultural University, Beijing, China.
authors.forename: Qian-Qian
authors.initials: QQ
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Agrobiotechnology and College of Biological Science, China Agricultural University, Beijing, China.
authors.forename: Jian-Wei
authors.initials: JW
authors.lastname: Li
authors.affiliation: State Key Laboratory of Agrobiotechnology and College of Biological Science, China Agricultural University, Beijing, China.
authors.forename: Yu-Mei
authors.initials: YM
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Agrobiotechnology and College of Biological Science, China Agricultural University, Beijing, China.
authors.forename: Xiao-Rong
authors.initials: XR
authors.lastname: An
authors.affiliation: State Key Laboratory of Agrobiotechnology and College of Biological Science, China Agricultural University, Beijing, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Hou
mesh_terms: D044503:5-Methylcytosine / Q000031:analogs & derivatives / Q000378:metabolism; D000818:Animals; D001205:Ascorbic Acid / Q000378:metabolism / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics*; D002460:Cell Line; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D049308:Dioxygenases; D018507:Gene Expression Regulation, Developmental; D055785:Gene Knockdown Techniques; D051379:Mice; D024510:Muscle Development / Q000235:genetics*; D032448:Myoblasts, Skeletal / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011518:Proto-Oncogene Proteins / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; D011518:Proto-Oncogene Proteins; C011865:5-hydroxymethylcytosine; D044503:5-Methylcytosine; D049308:Dioxygenases; C551914:Tet2 protein, mouse; D001205:Ascorbic Acid