RGD Reference Report - MicroRNA-96 regulates RGC-5 cell growth through caspase-dependent apoptosis. - Rat Genome Database

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MicroRNA-96 regulates RGC-5 cell growth through caspase-dependent apoptosis.

Authors: Wang, S  Li, K 
Citation: Wang S and Li K, Int J Clin Exp Med. 2014 Oct 15;7(10):3694-702. eCollection 2014.
RGD ID: 11553931
Pubmed: PMID:25419419   (View Abstract at PubMed)
PMCID: PMC4238503   (View Article at PubMed Central)

PURPOSE: This study is to investigate functional role of microRNA 96 (miR-96) in regulating cell survival and caspase-dependent apoptosis in rat retinal ganglion cell line RGC-5 cells. METHOD: RGC-5 cells were cultured in vitro. Lentiviral vectors of miR-96 inhibitor, miR-96 mimics and non-specific miRNA were applied. The effects of up-regulating or down-regulating miR-96 on cell survival, apoptosis and activation of apoptosis-related caspase proteins were assessed by MTT assay, TUNEL staining and western blotting analysis, respectively. Direct binding of miR-96 and its suspected target caspase protein, caspase 2 (Casp-2), was examined by luciferase reporter assay. Endogenous CASP2 gene was then silenced by siRNA in RGC-5 cells. The effect of down-regulating CASP2 on miR-96 mediated apoptosis in RGC-5 was assessed by western blotting and TUNEL staining. RESULTS: Lentivirus mediated- miR-96 up-regulation significantly reduced RGC-5 cell viability 12 hours after transfection, whereas miR-96 down-regulation had no effect on RGC-5 cell survival up to 72 hours. Up-regulating miR-96 induced significant apoptosis in RGC-5 cells, whereas down-regulating miR-96 had little effect on RGC-5 apoptosis. Western blotting analysis demonstrated that RGC-5 apoptosis induced by overexpressing miR-96 is associated with activation of caspase proteins, including caspase-2, 3, 9. Luciferase reporter assay confirmed that Casp-2 was directly bound by miR-96 in RGC-5 cells, and siRNA-regulated silencing of CASP2 gene rescued the apoptosis in RGC-5 induced by over-expressing miR-96. CONCLUSION: miR-96 is an important regulator on RGC-5 cell survival and apoptosis, possibly through Casp-2.

Gene Ontology Annotations    Click to see Annotation Detail View

Biological Process
TermQualifierEvidenceWithReferenceNotesSourceOriginal Reference(s)
positive regulation of neuron apoptotic process  IDA 11553931 RGD 

Objects Annotated

Genes (Rattus norvegicus)
Mir96  (microRNA 96)


Additional Information