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Published on: July 17, 2018
MicroRNA-214 protects L6 skeletal myoblasts against hydrogen peroxide-induced apoptosis
Wei Chen1, Ya-Nan Zhang1, Qiong-Qiong Jia2
1Department of Histology and Embryology, Hebei Medical University, Shijiazhuang, Hebei, China.
Abstract:
MicroRNAs (miRNAs) have been reported as key gene regulators, and they control many fundamental biological processes. Previously, we demonstrated that miR-214 had a protective effect against myocardial apoptosis and myocardial fibrosis. In this study, we sought to investigate the expression of miR-214 in L6 skeletal myoblast (SKM), the regulatory effect of miR-214 on hydrogen peroxide (H2O2) induced cell apoptosis and the underlying mechanisms of the antiapoptotic effect. MiR-214 expression was up-regulated by H2O2 in a dose and time-dependent manner in L6 SKMs. To investigate the regulatory effects of miR-214 on L6 SKM, both gain-of-function and loss-of-function approaches were applied. The results showed that miR-214 improved cell survival and inhibited cell apoptosis, and blockage of miR-214 abrogated the protective effect on cell survival and resistance to apoptosis. Phosphatase and tensin homolog (PTEN) was negatively regulated by miR-214, and PTEN inhibitor obviously reversed the effect of miR-214 blockage on enhancing cell apoptosis. In addition, miR-214 up-regulated antiapoptotic protein Bcl-2, down-regulated proapoptotic protein Bax, prevented release of cytochrome c and inhibited caspase-3 activation. In summary, H2O2-induced injury increases miR-214 expression in L6 SKM, and miR-214 contributes to the protection of L6 SKM against apoptosis via lowering PTEN and subsequently inhibiting the mitochondrial-mediated caspase-dependent apoptotic signaling pathway.
Insights
Hydrogen peroxide (H2O2) increases microRNA-214 (miR-214) in skeletal muscle cells. This miR-214 protects cells from apoptosis by regulating PTEN and downstream signaling pathways.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- MicroRNAs (miRNAs) are key gene regulators involved in fundamental biological processes.
- Previous studies indicated miR-214 protects against myocardial apoptosis and fibrosis.
- Oxidative stress is implicated in skeletal muscle dysfunction.
Purpose of the Study:
- To investigate miR-214 expression in L6 skeletal myoblasts (SKMs) under H2O2 exposure.
- To determine the regulatory role of miR-214 in H2O2-induced SKM apoptosis.
- To elucidate the underlying antiapoptotic mechanisms of miR-214.
Main Methods:
- L6 SKMs were treated with varying doses and durations of H2O2.
- Gain-of-function and loss-of-function approaches were used to manipulate miR-214 levels.
- Cell survival, apoptosis, PTEN expression, and key apoptotic pathway proteins (Bcl-2, Bax, cytochrome c, caspase-3) were assessed.
Main Results:
- H2O2 dose- and time-dependently upregulated miR-214 expression in L6 SKMs.
- Overexpression of miR-214 enhanced cell survival and inhibited apoptosis.
- Inhibition of miR-214 promoted apoptosis, which was partially reversed by a PTEN inhibitor.
- miR-214 downregulated PTEN, upregulated Bcl-2, downregulated Bax, prevented cytochrome c release, and inhibited caspase-3 activation.
Conclusions:
- H2O2-induced injury elevates miR-214 expression in L6 SKMs.
- miR-214 confers protection against apoptosis in L6 SKMs.
- The protective mechanism involves PTEN downregulation and subsequent inhibition of the mitochondrial-mediated, caspase-dependent apoptotic pathway.
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