Journal
CELLULAR PHYSIOLOGY AND BIOCHEMISTRY
Volume 46, Issue 1, Pages 351-364Publisher
KARGER
DOI: 10.1159/000488435
Keywords
Cam kinase IV (CaMKIV); C2C12 cell; Gene knockdown; IFN-gamma
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Funding
- National Natural Science Foundation of China [81371924, 81572102]
- National Key Research and Development Plan [2017YFC1105003]
- GuangDong Provincial Natural Science Foundation [2014A030313276]
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Background/Aims: Whether calcium/calmodulin-dependent protein kinase IV (CaMKIV) plays a role in regulating immunologic features of muscle cells in inflammatory environment, as it does for immune cells, remains mostly unknown. In this study, we investigated the influence of endogenous CaMKIV on the immunological characteristics of myoblasts and myotubes received IFN-gamma stimulation. Methods: C2C12 and murine myogenic precursor cells (MPCs) were cultured and differentiated in vitro, in the presence of pro-inflammatory IFN-gamma. CaMKIV shRNA lentivirus transfection was performed to knockdown CaMKIV gene in C2C12 cells. pEGFP-N1-CaMKIV plasmid was delivered into knockout cells for recovering intracellular CaMKIV gene level. CREB1 antagonist KG-501 was used to block CREB signal. qPCR, immunoblot analysis, or immunofluorescence was used to detect mRNA and protein levels of CaMKIV, immuno-molecules, or pro-inflammatory cytokines and chemokines. Costimulatory molecules expression was assessed by FACS analysis. Results: IFN-gamma induces the expression or up-regulation of MHC-I/II and TLR3, and the up-regulation of CaMKIV level in muscle cells. In contrast, CaMKIV knockdown in myoblasts and myotubes leads to expression inhibition of the above immuno-molecules. As well, CaMKIV knockdown selectively inhibits pro-inflammatory cytokines/chemokines, and co-stimulatory molecules expression in IFN-gamma treated myoblasts and myotubes. Finally, CaMKIV knockdown abolishes IFN-gamma induced CREB pathway molecules accumulation in differentiated myotubes. Conclusions: CaMKIV can be induced to up-regulate in muscle cells under inflammatory condition, and positively mediates intrinsic immune behaviors of muscle cells triggered by IFN-gamma. (C) 2018 The Author(s) Published by S. Karger AG, Basel
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