Abstract：
BACKGROUND:We intended to examine the functional role of microRNA 26 (miR-26a) in regulating H2O2-induced cytotoxicity and apoptosis in RGC-5 cells in vitro. METHOD:Various concentrations of H2O2 (0-1000 μM) were added in RGC-5 culture. Cell cytotoxicity was monitored by viability assay and gene expression level of miR-26a examined by qRT-PCR. MicroRNA-26a mimic was then applied in the RGC-5 culture to examine its effect on upregulating endogenous miR-26a and rescuing H2O2-induced cytotoxicity. TUNEL immunostaining assay was used to further assess the protective effect of upregulating miR-26a on H2O2-induced apoptosis in RGC-5 cells. Direct targeting of miR-26a on Phosphatase and tensin homolog (PTEN) signaling pathway was assessed by luciferase assay and western blotting. PTEN was then ectopically over-expressed in RGC-5. And its effects on miR-26a mediated apoptosis protection in RGC-5 were investigated by western blot and TUNEL assay. RESULTS:H2O2 induced cytotoxicity and down-regulated miR-26a in dose-dependent manner in RGC-5 cells. MiR-26a-mimic upregulated endogenous miR-26a gene levels, and then reduced H2O2-induced cytotoxicity, as well as H2O2-induced apoptosis in RGC-5 cells. PTEN was directly targeted by miR-26a. PTEN protein was upregulated, and phosphorylated AKT protein down-regulated while miR-26a was upregulated to reduce H2O2-induced apoptosis. Finally, overexpressing PTEN reversed the protective effect of miR-26a upregulation on RGC-5 apoptosis. CONCLUSION:Upregulating miR-26a protects RGC-5 cell against cytotoxicity and apoptosis, probably through down-regulation of PTEN.
journal_name
Biochem Biophys Res Communjournal_title
Biochemical and biophysical research communicationsauthors
Kang Y,Jia P,Zhao H,Hu C,Yang Xdoi
10.1016/j.bbrc.2015.02.164subject
Has Abstractpub_date
2015-05-01 00:00:00pages
164-9issue
2eissn
0006-291Xissn
1090-2104pii
S0006-291X(15)00411-8journal_volume
460pub_type
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