Milk fat globule membrane protein promotes C2C12 cell proliferation through the PI3K/Akt signaling pathway

Li, He; Xu, Weili; Ma, Ying; Zhou, Shaobo; Xiao, Ran

doi:10.1016/j.ijbiomac.2018.04.026

Cited by 25 publications

(30 citation statements)

References 48 publications

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“…7 was shown that MFG-E8 can up-regulate the expression of phosphorylation of PI3K and ERK. Wortmannin plays a central role to negatively regulate C2C12 cell by inhibiting PI3K/Akt-mediated PI3 phosphorylation and activation of the PI3K/Akt pathway [9,27]. Furthermore, PI3K and ERK phosphorylation was reduced by combination treatment with wortmannin and U0126, U0126 exerted effects similar to wortmannin on ERK phosphorylation either alone or in combination with wortmannin (p<0.05).…”

Section: Verification Of Phosphoproteins By Western Blottingmentioning

confidence: 90%

“…Our previous research found that MFG-E8 promoted C2C12 cells proliferation via PI3K/Akt signal pathway, which can activate P70S6k through phosphorylation of Akt. Therefore, the results was further to prove that MFG-E8 promote C2C12 cell proliferation mainly via PI3K/Akt signal pathway [9].…”

Section: The Up-regulation Of Myoblast C2c12 Cell Proteinmentioning

confidence: 92%

“…The protein concentration was quantified using the BCA kit. 100 μg protein was loaded onto a 1-D SDS gel (12% polyacrylamide) [9]. The gel was stained with Coomassie brilliant blue R-250.…”

Section: Sds-pagementioning

confidence: 99%

“…The protein in C2C12 cell components were further analysed with capillary LC-MS/MS based on our previous research [9]. The identification and quantification of protein components in C2C12 cells was confirmed using LC-MS/MS.…”

Section: Liquid Chromatography (Lc)-electrospray Ionization (Esi) Tanmentioning

confidence: 99%

“…PI3K also plays a central role in cell signaling and leads to cell proliferation, survival, motility, secretion, and specialized cell responses, such as the respiratory burst of granulocytes [9,27]. Based on the KEGG analysis, the mechanism of MFG-E8 and the relationship between the PI3K/Akt and MAPK/ERK signal pathway.…”

Section: Verification Of Phosphoproteins By Western Blottingmentioning

confidence: 99%

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

Guan

et al. 2019

International Journal of Biological Macromolecules

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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.

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Section: Verification Of Phosphoproteins By Western Blottingmentioning

confidence: 90%

Section: The Up-regulation Of Myoblast C2c12 Cell Proteinmentioning

confidence: 92%

Section: Sds-pagementioning

confidence: 99%

Section: Liquid Chromatography (Lc)-electrospray Ionization (Esi) Tanmentioning

confidence: 99%

Section: Verification Of Phosphoproteins By Western Blottingmentioning

confidence: 99%

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

Guan

et al. 2019

International Journal of Biological Macromolecules

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Melatonin inhibits embryonic rat H9c2 cells growth through induction of apoptosis and cell cycle arrest via PI3K‐AKT signaling pathway

Zhao

Xia

et al. 2021

Birth Defects Research

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Background Our recent epidemiological study revealed that maternal sleep during the periconceptional period should be involved in the risk of congenital heart disease (CHD) in offspring. Melatonin, a sleep related hormone, has been suggested to play a crucial role in embryonic development based on the emerging evidence. In this study, we set out to assess the effect of melatonin on the embryonic cardiac cell growth and to explore the underlying mechanisms. Methods We observed the effect of different gradient doses of melatonin as 10, 100, or 1,000 μM on cell proliferation in H9c2 embryonic rat cardiac cells. Furthermore, flow cytometry was applied to evaluate the impact on apoptosis and cell cycle. RNA‐seq was conducted to screen the changes in expression of mRNA and signaling pathways. Quantitative Real‐Time‐PCR (qRT‐PCR) was then conducted to validate the results. Results It was observed that melatonin could inhibit H9c2 cell growth, at the doses of 100 and 1,000 μM, but not at 10 μM. Moreover, melatonin ranged from 100 to 1,000 μM could instigate cell cycle arrest at G1 phase and simulate apoptosis, in a dose‐dependent manner. In addition, melatonin was found to down‐regulate the expression of a number of genes, which are related to heart development (SPARC, IFITM3, TNNT2, LOX), and PI3K‐Akt signaling pathway activation (FN1, HSP90B1, THBS1, MFGE8, and CLU). Conclusions Our findings suggested that high level of melatonin could be capable of inhibiting growth through the induction of apoptosis and cell cycle arrest via PI3K‐AKT signaling pathway, thereby interfering with embryonic heart development. Considering this study is based on H9c2 embryonic rat cardiac cells, future additional studies using human embryonic cardiac cell are warranted.

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An image‐based Abplex method for high‐throughput GPCRs antibody discovery

Jiang,

Wang,

et al. 2023

Biotechnology Journal

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As the field of antibody therapeutics advances rapidly, membrane proteins, particularly G protein‐coupled receptors (GPCRs), have emerged as highly sought‐after drug targets. However, the challenges associated with extracting membrane proteins have created a demand for effective antibody screening systems targeting these proteins. In this study, we propose developing an innovative antibody screening strategy (Abplex) based on high‐content imaging. This approach leverages intact cells that express target membrane proteins, facilitating the presentation of proteins in their native conformation. Furthermore, it acquires both specific and non‐specific binding signals in a single well, thereby bolstering the robustness of the outcomes. The technique involves just one step and can be completed within 50 min, enabling the analysis of a single sample in just one second. The amalgamation of dependable experimental findings, a simplified workflow, reduced hands‐on time, and a swift analytical pace positions our method for superior throughput and precision when juxtaposed with traditional techniques such as CbELISA and FACS. Moreover, we introduce the concept of cell barcoding, wherein cells are labeled with different fluorescence spatial patterns. This feature allows for multiplexed detection to meet the needs of various experiments. The characteristics of Abplex promise to expedite GPCR‐targeting antibody discovery, advance therapeutics and enable new disease treatments.

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Milk fat globule membrane protein promotes C2C12 cell proliferation through the PI3K/Akt signaling pathway

Cited by 25 publications

References 48 publications

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

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

Melatonin inhibits embryonic rat H9c2 cells growth through induction of apoptosis and cell cycle arrest via PI3K‐AKT signaling pathway

An image‐based Abplex method for high‐throughput GPCRs antibody discovery

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