Many intracellular signaling pathways regulate skeletal muscle differentiation. Among them, PI3K/AKT pathway plays an important role. But the mechanisms of chromatin regulation remain unclear. In this study, the murine C2C12 myoblast cell line was used to investigate the expression of Myogenin and MCK genes during the skeletal muscle differentiation. Western blotting analysis showed that the expression of Myogenin and MCK protein was increased signifi-cantly after PI3K/AKT activator treatment for 24 h during the C2C12 cell differentiation and the expression of H3K27me3 demethylase UTX was also increased. Chromatin immunoprecipitation (ChIP) and quantitative PCR (Q-PCR) analysis showed that the enrichment of H3K27me3 on the promoter regions of Myogenin and MCK genes and the enhancer region of MCK gene were decreased. It was opposite to the PI3K/AKT inhibitor treatment. We concluded that the PI3K/AKT pathway maybe regulate skeletal muscle differentiation by regulating the expression of UTX gene to change the enrichment of H3K27me3 on the target genes.
LI Jing ZHANG Yun-Sheng LI Ning HU Xiao-Xiang SHI Guo-Qing LIU Shou-Ren LIU Nan
. Expression of Myogenin and MCK genes regulated by PI3K/AKT pathway[J]. Hereditas(Beijing), 2013
, 35(5)
: 637
-642
.
DOI: 10.3724/SP.J.1005.2013.00637
[1] Zetser A, Gredinger E, Bengal E. p38 mitogen-activated protein kinase pathway promotes skeletal muscle differentiation: Participation of the mef2c transcription factor. Biol Chem, 1999, 274(8): 5193-5200.
[2] Himeda CL, Tai PW, Hauschka SD. Analysis of muscle gene transcription in cultured skeletal muscle cells. Methods Mol Biol, 2012, 798(7): 425-443.
[3] Figueroa A, Cuadrado A, Fan JS, Atasoy U, Muscat GE, Muñoz-Canoves P, Gorospe M, Muñoz A. Role of HuR in Skeletal myogenesis through coordinate regulation of muscle differentiation genes. Mol Cell Biol, 2003, 23(14): 4991-5004.
[4] Kaneko S, Feldman RI, Yu L, Wu ZG, Gritsko T, Shelley SA, Nicosia SV, Nobori T, Cheng JQ. Positive feedback regulation between Akt2 and MyoD during muscle differentiation: Cloning of Akt2 promoter. J Biol Chem, 2002, 277(26): 23230-23235.
[5] Serra C, Palacios D, Mozzetta C, Forcales SV, Morantte I, Ripani M, Jones DR, Du KY, Jhala US, Simone C, Puri PL. Functional interdependence at the chromatin level between the MKK6/p38 and IGF1/PI3K/AKT pathways during muscle differentiation. Mol Cell, 2007, 28(2): 200-213.
[6] Seenundun S, Rampalli S, Liu QC, Aziz A, Palii C, Hong S, Blais A, Brand M, Ge K, Dilworth FJ. UTX mediates demethylation of H3K27me3 at muscle-specific genes during myogenesis. EMBO J, 2010, 29(8): 1401-1411.
[7] 郭芬, 刘兆宇, 李月琴, 李弘剑, 周天鸿. Prosaposin对细胞增殖和凋亡的调控及其分子机制. 遗传, 2009, 31(12): 1226-1232.
[8] 郭云学, 莫德林, 陈瑶生, 张悦, 张云, 肖书奇, 刘小红. PI3K/AKT抑制剂渥曼青霉素对猪前体脂肪细胞增殖和凋亡的影响. 遗传, 2012, 34(10): 1282-1290.
[9] Jiang BH, Aoki M, Zheng JZ, Li J, Vogt PK. Myogenic signaling of phosphatidylinositol 3-kinase requires the serine-threonine kinase Akt/protein kinase B. Proc Natl Acad Sci USA, 1999, 96(5): 2077-2081.
[10] Rommel C, Bodine SC, Clarke BA, Rossman R, Nunez L, Stitt TN, Yancopoulos GD, Glass DJ. Mediation of IGF-1-induced skeletal myotube hypertrophy by PI(3)K/Akt/mTOR and PI(3)K/Akt/GSK3 pathways. Nature Cell Biol, 2001, 3(11): 1009-1013.
[11] Fuks F. DNA methylation and histone modification: teaming up to silence genes. Curr Opin Genet Dev, 2005, 15(5): 490-495.
[12] Bernstein BE, Meissner A, Lander ES. The mammalian epigenome. Cell, 2007, 128(4): 669-681.
[13] Cha TL, Zhou BP, Xia WY, Wu YD, Yang CC, Chen CT, Ping B, Otte AP, Hung MC. AKT-mediated phosphorylation of EZH2 suppresses methylation of lysine 27 in histone H3. Science, 2005, 310(5746): 306-310.
[14] Wang JK, Tsai MC, Poulin G, Adler AS, Chen S, Liu H, Shi Y, Chang HY. The histone demethylase UTX enables RB-dependent cell fate control. Genes Dev, 2010, 24(4): 327-332.