To reveal the microRNA (miRNA) expression profile and related roles in rat HSCs during activation.
miRNA expression profiling was analyzed in quiescent and in culture-activated HSCs by microarray. The differentially expressed miRNAs, as verified by RT-PCR, were subjected to gene ontology (GO) analysis. Furthermore, the effects of miR-16 and miR-15b on the apoptosis of activated HSCs were investigated by Hoechst 33258, TUNEL staining and annexin-V/PI labeling flow cytometry. The underlying mechanism related to Bcl-2 and caspases was assessed.
The upregulated and downregulated miRNAs in activated HSCs were 12 miRNAs and 9 miRNAs, respectively. The differential expression of miR-16, -15b, -122, -138, -143, and -140 was validated. High-enrichment GOs containing apoptosis-related targeted genes and miRNA–gene networks characterized by Bcl-2, which was targeted by the miR-15/16 family, uncovered the critical role of miR-16 and miR-15b in apoptosis. Restoring the intracellular miRNAs by miR-16 and miR-15b administration greatly reduced Bcl-2, and increased the expression of caspases 3, 8, and 9. Significantly elevated rates of apoptosis were then induced in activated HSCs.
The activation of HSCs relate to 21 miRNAs. Among these, mir-15b and miR-16 may be essential for apoptosis by targeting Bcl-2 and the caspase signaling pathway.
Abbreviations:miRNA (microRNA), HSC (hepatic stellate cell), GO (gene ontology), ECM (extracellular matrix), 3′ UTR (3′ untranslated region), ALB (albumin), CK-19 (cytokeratin-19), FDR (false discovery rate), SD (standard deviation)
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Published online: February 02, 2009
Accepted: November 6, 2008
Received in revised form: October 28, 2008
Received: July 24, 2008Associate Editor: C. Trautwein
☆The authors declare that they do not have anything to disclose regarding funding from industries or conflict of interest with respect to this paper.
© 2009 European Association for the Study of the Liver. Published by Elsevier Inc. All rights reserved.