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Overexpression of miR-483-5p/3p cooperate to inhibit mouse liver fibrosis by suppressing the TGF-β stimulated HSCs in transgenic mice.

Li F, Ma N, Zhao R, Wu G, Zhang Y, Qiao Y, Han D, Xu Y, Xiang Y, Yan B, Jin J, Lv G, Wang L, Xu C, Gao X, Luo S - J. Cell. Mol. Med. (2014)

Bottom Line: MicroRNAs (miRNAs) are recently discovered molecules that regulate the expression of genes involved in liver disease.Many reports demonstrate that miR-483-5p and miR-483-3p, which originate from miR-483, are up-regulated in HCC, and their oncogenic targets have been identified.We demonstrate that miR-483-5p/3p acts together to target two pro-fibrosis factors, platelet-derived growth factor-β and tissue inhibitor of metalloproteinase 2, which suppress the activation of hepatic stellate cells (HSC) LX-2.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Harbin Medical University, Harbin, China.

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miR-483-5p and miR-483-3p inhibit transforming growth factor-β (TGF-β) stimulated LX-2 cells. (A) Expression of α-SMA at the transcriptional level in quiescent versus TGF-β stimulated LX-2 cells. (B) Expression of α-SMA at the translational level in quiescent versus TGF-β stimulated LX-2 cells. (C) qRT-PCR analysis for miR-483-5p and miR-483-3p was performed with RNA extracts from quiescent and activated hepatic stellate cells (n = 4). (D) Regulation of α-SMA proteins after miR-483 transfection. (E) Immunofluorescence for α-SMA proteins after miR-483 transfection. Three independent experiments were performed; **P < 0.01.
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fig03: miR-483-5p and miR-483-3p inhibit transforming growth factor-β (TGF-β) stimulated LX-2 cells. (A) Expression of α-SMA at the transcriptional level in quiescent versus TGF-β stimulated LX-2 cells. (B) Expression of α-SMA at the translational level in quiescent versus TGF-β stimulated LX-2 cells. (C) qRT-PCR analysis for miR-483-5p and miR-483-3p was performed with RNA extracts from quiescent and activated hepatic stellate cells (n = 4). (D) Regulation of α-SMA proteins after miR-483 transfection. (E) Immunofluorescence for α-SMA proteins after miR-483 transfection. Three independent experiments were performed; **P < 0.01.

Mentions: The activation of HSCs is a key process during liver fibrosis in vivo. Because we observed the inhibition of α-SMA in miR-483 transgenic mice, we investigated the role of miR-483 in the activation of HSCs. First, stimulation of the human HSC cell line LX-2 with recombinant TGF-β, a major cytokine involved in HSC activation (Fig.3A and B), led to an increase in TIMP2 and PDGF-β (Fig. S2A) and significant decrease in miR-483 expression (Fig.3C). These results are consistent with the results observed for CCl4-induced liver fibrosis in transgenic mice. We then transfected either miR-483 or a control sequence into the TGF-β stimulated LX-2 cells. α-SMA was down-regulated by overexpression of miR-483 at the translational level (Fig.3D). These results are consistent with the role of miR-483 in vivo. Our data suggest that overexpression of miR-483 regulates liver fibrosis by inhibiting the activation of HSCs in transgenic mice. Next, we identified the targets of miR-483 in this regulatory process.


Overexpression of miR-483-5p/3p cooperate to inhibit mouse liver fibrosis by suppressing the TGF-β stimulated HSCs in transgenic mice.

Li F, Ma N, Zhao R, Wu G, Zhang Y, Qiao Y, Han D, Xu Y, Xiang Y, Yan B, Jin J, Lv G, Wang L, Xu C, Gao X, Luo S - J. Cell. Mol. Med. (2014)

miR-483-5p and miR-483-3p inhibit transforming growth factor-β (TGF-β) stimulated LX-2 cells. (A) Expression of α-SMA at the transcriptional level in quiescent versus TGF-β stimulated LX-2 cells. (B) Expression of α-SMA at the translational level in quiescent versus TGF-β stimulated LX-2 cells. (C) qRT-PCR analysis for miR-483-5p and miR-483-3p was performed with RNA extracts from quiescent and activated hepatic stellate cells (n = 4). (D) Regulation of α-SMA proteins after miR-483 transfection. (E) Immunofluorescence for α-SMA proteins after miR-483 transfection. Three independent experiments were performed; **P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4508137&req=5

fig03: miR-483-5p and miR-483-3p inhibit transforming growth factor-β (TGF-β) stimulated LX-2 cells. (A) Expression of α-SMA at the transcriptional level in quiescent versus TGF-β stimulated LX-2 cells. (B) Expression of α-SMA at the translational level in quiescent versus TGF-β stimulated LX-2 cells. (C) qRT-PCR analysis for miR-483-5p and miR-483-3p was performed with RNA extracts from quiescent and activated hepatic stellate cells (n = 4). (D) Regulation of α-SMA proteins after miR-483 transfection. (E) Immunofluorescence for α-SMA proteins after miR-483 transfection. Three independent experiments were performed; **P < 0.01.
Mentions: The activation of HSCs is a key process during liver fibrosis in vivo. Because we observed the inhibition of α-SMA in miR-483 transgenic mice, we investigated the role of miR-483 in the activation of HSCs. First, stimulation of the human HSC cell line LX-2 with recombinant TGF-β, a major cytokine involved in HSC activation (Fig.3A and B), led to an increase in TIMP2 and PDGF-β (Fig. S2A) and significant decrease in miR-483 expression (Fig.3C). These results are consistent with the results observed for CCl4-induced liver fibrosis in transgenic mice. We then transfected either miR-483 or a control sequence into the TGF-β stimulated LX-2 cells. α-SMA was down-regulated by overexpression of miR-483 at the translational level (Fig.3D). These results are consistent with the role of miR-483 in vivo. Our data suggest that overexpression of miR-483 regulates liver fibrosis by inhibiting the activation of HSCs in transgenic mice. Next, we identified the targets of miR-483 in this regulatory process.

Bottom Line: MicroRNAs (miRNAs) are recently discovered molecules that regulate the expression of genes involved in liver disease.Many reports demonstrate that miR-483-5p and miR-483-3p, which originate from miR-483, are up-regulated in HCC, and their oncogenic targets have been identified.We demonstrate that miR-483-5p/3p acts together to target two pro-fibrosis factors, platelet-derived growth factor-β and tissue inhibitor of metalloproteinase 2, which suppress the activation of hepatic stellate cells (HSC) LX-2.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Harbin Medical University, Harbin, China.

Show MeSH
Related in: MedlinePlus