title: Reversine induces multipotency of lineage-committed cells through epigenetic silencing of miR-133a.
abstract: Reversine has been shown to induce dedifferentiation of C2C12 murine myoblasts into multipotent progenitor cells. However, little is known about the key regulators mediating the dedifferentiation induced by reversine. Here, we show that large scale miRNA gene expression profiling of reversine-treated C2C12 myoblasts identifies a down-regulated miRNA, miR-133a, involved in dedifferentiation of myoblasts. Reversine treatment results in up- and down-regulated miRNA profiles. Among miRNAs affected by reversine, the level of muscle-specific miR-133a, which has been shown to be up-regulated during muscle development and to suppress differentiation into other lineages, is markedly reduced by treatment of C2C12 myoblasts with reversine. In parallel, reversine decreases the expression and recruitment of myogenic factor, SRF, to the enhancer regions of miR-133a. Sequentially, down-regulation of miR-133a by reversine is accompanied by a decrease in active histone modifications including trimethylation of histone H3K4 and H3K36, phosphorylation of H3S10, and acetylation of H3K14 on the miR-133a promoter, leading to dissociation of RNA polymerase II from the promoter. Furthermore, inhibition of miR-133a by transfection of C2C12 myoblasts with miR-133a inhibitor increases the expression of osteogenic lineage marker, Ogn, and adipotenic lineage marker, ApoE, similar to that in response to reversine. In contrast, the co-overexpression of miR-133a mimic reversed the effect of reversine on C2C12 myoblast dedifferentiation. Taken together, the results indicate that reversine induces a multipotency of C2C12 myoblasts by suppression of miR-133a expression through depletion of active histone modifications, and suggest that miR-133a is a potential miRNA regulating the reversine-induced dedifferentiation. Collectively, our findings provide a mechanistic rationale for the application of reversine to dedifferentiation of somatic cells.
authors: Research Center for Epigenome Regulation, School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors: Munkyung
authors: M
authors: Kim
authors: Research Center for Epigenome Regulation, School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors: Sang Ah
authors: SA
authors: Yi
authors: Research Center for Epigenome Regulation, School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors: Hyunwoo
authors: H
authors: Lee
authors: Research Center for Epigenome Regulation, School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors: So Young
authors: SY
authors: Bang
authors: Research Center for Epigenome Regulation, School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors: Eun Kyung
authors: EK
authors: Park
authors: Research Center for Epigenome Regulation, School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors: Min Gyu
authors: MG
authors: Lee
authors: Research Center for Epigenome Regulation, School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors: Ki Hong
authors: KH
authors: Nam
authors: Research Center for Epigenome Regulation, School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors: Ji Hee
authors: JH
authors: Yoo
authors: College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University, Incheon 406-840, Republic of Korea.
authors: Dong Hoon
authors: DH
authors: Lee
authors: College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University, Incheon 406-840, Republic of Korea.
authors: Hyun-Wook
authors: HW
authors: Ryu
authors: College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University, Incheon 406-840, Republic of Korea. Electronic address: soheekwon@yonsei.ac.kr.
authors: So Hee
authors: SH
authors: Kwon
authors: Research Center for Epigenome Regulation, School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea. Electronic address: jhhan551@skku.edu.
authors: Jeung-Whan
authors: JW
authors: Han
mesh_terms: D000107:Acetylation / Q000187:drug effects; D000818:Animals; D015153:Blotting, Western; D054337:Cell Dedifferentiation / Q000187:drug effects / Q000235:genetics; D002460:Cell Line; D019070:Cell Lineage / Q000187:drug effects / Q000235:genetics; D044127:Epigenesis, Genetic / Q000187:drug effects*; D020869:Gene Expression Profiling; D020868:Gene Silencing / Q000187:drug effects*; D006657:Histones / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D008745:Methylation / Q000187:drug effects; D051379:Mice; D035683:MicroRNAs / Q000235:genetics*; D009025:Morpholines / Q000494:pharmacology*; D039902:Multipotent Stem Cells / Q000187:drug effects* / Q000378:metabolism; D032446:Myoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D010766:Phosphorylation / Q000187:drug effects; D011401:Promoter Regions, Genetic / Q000235:genetics; D011687:Purines / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D026362:Serum Response Factor / Q000235:genetics / Q000378:metabolism
chemical_list: D006657:Histones; D036341:Intercellular Signaling Peptides and Proteins; D035683:MicroRNAs; C517732:Mirn133 microRNA, mouse; D009025:Morpholines; C512906:Ogn protein, mouse; D011687:Purines; D026362:Serum Response Factor; C484369:2-(4-morpholinoanilino)-6-cyclohexylaminopurine
keywords: Dedifferentiation; Histone modifications; Myogenesis; Reversine; miR-133a