MFG-E8 induced differences in proteomic profiles in mouse C2C12 cells and its effect on PI3K/Akt and ERK signal pathways

Abstract

Milk fat globule-EGF factor 8 (MFG-E8) is one of the major proteins in milk fat globule membrane. In this study, mouse-derived C2C12 myoblast cells were served as an experimentally tractable model system for investigating the molecular basis of skeletal muscle cell specification and development. To examine the biochemical adaptations associated with myocytes formation comprehensively, a liquid chromatography coupled with tandem mass spectrometry label-free semi-quantitative approach was used to analyse the myogenic C2C12 proliferation program. Over 1987 proteins were identified in C2C12 cells. The MFG-E8 (200 g/mL) and MFG-E8 (500 g/mL) with significant differences were compared based on the relative abundance. The result profiles of regulation of MFG-E8 to the expression of proteins in C2C12 cells revealed that differential waves of expression of proteins linked to intracellular signaling, transcription, cytoarchitecture, adhesion, metabolism, and muscle contraction across during the C2C12 cell proliferation process. Based on the analysis of KEGG and STRING database, further to verification the expression of PI3K and ERK phosphorylation levels by Western blot. This study found that the data of proteomic was complementary to recent MFG-E8 studies of protein expression patterns in developing myotubes and provided a holistic framework for understanding how diverse biochemical processes are coordinated at the cellular level during skeletal muscle development.

Bovine Milk Fat Globule Epidermal Growth Factor VIII activates PI3K/Akt signaling pathway and attenuates sarcopenia in rat model induced by d-galactose

“…The MFG-E8 was extracted using an electric cream separator, and separated according to cellulose DEAE-52 ion exchange chromatography method. Our previous work has demonstrated that MFG-E8 can promote cell proliferation and differentiation via the PI3K/Akt and ERK signal pathway in the in vitro C2C12 cells (Li et al 2019). In this study, we focused on the in vivo rat experiment to investigate the effect of MFG-E8 on age-related sarcopenia.…”

“…We have previously demonstrated in vitro C2C12 cells that MFG-E8 could promote cell proliferation and differentiation (He et al 2017;Li et al 2017). Based on the results of cell differential proteomics, we clarified that MFG-E8 promoting cell proliferation is related to 60S ribosomal protein L29, Hsp 40, integrin α-V, serine/threonine protein phosphatase 2A and insulin degrading enzyme expression, mediating PI3K, MAPK, AMPK and Oxidative phosphorylation signaling pathways cascade, and PI3K plays a major role (Li et al 2019). In vitro experiment preliminarily demonstrated the possible benefits of MFG-E8 on sarcopenia (Li et al 2018).…”

The endocrine and epigenetic impact of persistent cow milk consumption on prostate carcinogenesis

“…Milk fat globule-EGF factor 8 MFG membrane proteins, predominantly MFG epidermal growth factor 8 (MFG-E8, also known as lactadherin), stimulates cell proliferation through the PI3K/AKT/mTORC1 signaling pathway [324][325][326][327] . Increased levels of MFG-E8 found in tissue and plasma exosomes from PCa patients have been compared with controls [327,328] .…”

Lifetime Impact of Cow’s Milk on Overactivation of mTORC1: From Fetal to Childhood Overgrowth, Acne, Diabetes, Cancers, and Neurodegeneration

“…Recent evidence indicates that protein palmitoylation is potentially involved in palmitate-induced mTORC1 activation, whereas 2-bromopalmitate, a protein palmitoylation inhibitor, ameliorated palmitate-triggered mTORC1 activation [110]. Furthermore, MFG membrane proteins, predominantly MFG-E8, promote cell proliferation through the PI3K/AKT/mTORC1 signaling pathway [111,112].…”