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Mir-34a is upregulated during liver regeneration in rats and is associated with the suppression of hepatocyte proliferation.

Chen H, Sun Y, Dong R, Yang S, Pan C, Xiang D, Miao M, Jiao B - PLoS ONE (2011)

Bottom Line: In BRL-3A cells, INHBB was identified as a direct target of miR-34a by luciferase reporter assay.A decrease of INHBB and Met was detected in regenerating liver.MiR-34a expression was upregulated during the late phase of liver regeneration.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Second Military Medical University, Shanghai, China.

ABSTRACT

Background: MicroRNAs are a class of small regulatory RNAs that modulate a variety of biological processes, including cellular differentiation, apoptosis, metabolism and proliferation. This study aims to explore the effect of miR-34a in hepatocyte proliferation and its potential role in liver regeneration termination.

Methodology/principal finding: MiR-34a was highly induced after partial hepatectomy. Overexpression of miR-34a in BRL-3A cells could significantly inhibit cell proliferation and down-regulate the expression of inhibin βB (INHBB) and Met. In BRL-3A cells, INHBB was identified as a direct target of miR-34a by luciferase reporter assay. More importantly, INHBB siRNA significantly repressed cell proliferation. A decrease of INHBB and Met was detected in regenerating liver.

Conclusion/significance: MiR-34a expression was upregulated during the late phase of liver regeneration. MiR-34a-mediated regulation of INHBB and Met may collectively contribute to the suppression of hepatocyte proliferation.

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Related in: MedlinePlus

MiR-34a induced growth inhibition in rat hepatocytes.(A) BRL-3A cells treated with miR-34a mimics (miR-34a) or negative control (NC) were seeded in 96-well plates and examined at indicated time points. The absorbance of methylthiazoletetrazolium by each sample was recorded at 570 nm after staining. (B,C) Cells treated with miR-34a or NC were analyzed by flow cytometry as described in “Materials and methods” for cell cycle distribution analysis.
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pone-0020238-g002: MiR-34a induced growth inhibition in rat hepatocytes.(A) BRL-3A cells treated with miR-34a mimics (miR-34a) or negative control (NC) were seeded in 96-well plates and examined at indicated time points. The absorbance of methylthiazoletetrazolium by each sample was recorded at 570 nm after staining. (B,C) Cells treated with miR-34a or NC were analyzed by flow cytometry as described in “Materials and methods” for cell cycle distribution analysis.

Mentions: To evaluate the effect of miR-34a in regulating rat liver cell proliferation, a MTT cell proliferation assay and a cell cycle analysis were used (described in “Materials and methods”). Briefly, BRL-3A immortalized rat liver cells were transfected with miR-34a or NC. MTT assay was performed at 2-day intervals, while cell cycle analysis was conducted 48 h after infection. As shown in Figure 2A, miR-34a markedly reduced BRL-3A cell growth at 4 d and more remarkably at 6 d (P<0.01). The growth inhibitory effect of miR-34a can also be sustained by the data of cell cycle analysis, in which the percentages of G2 phase cells in miR-34a and NC groups were (23.14±4.26)% and (8.48±2.93)% respectively, indicating a subpopulation of cells arrested in G2 phase by miR-34a (Figure 2B,C).


Mir-34a is upregulated during liver regeneration in rats and is associated with the suppression of hepatocyte proliferation.

Chen H, Sun Y, Dong R, Yang S, Pan C, Xiang D, Miao M, Jiao B - PLoS ONE (2011)

MiR-34a induced growth inhibition in rat hepatocytes.(A) BRL-3A cells treated with miR-34a mimics (miR-34a) or negative control (NC) were seeded in 96-well plates and examined at indicated time points. The absorbance of methylthiazoletetrazolium by each sample was recorded at 570 nm after staining. (B,C) Cells treated with miR-34a or NC were analyzed by flow cytometry as described in “Materials and methods” for cell cycle distribution analysis.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3105003&req=5

pone-0020238-g002: MiR-34a induced growth inhibition in rat hepatocytes.(A) BRL-3A cells treated with miR-34a mimics (miR-34a) or negative control (NC) were seeded in 96-well plates and examined at indicated time points. The absorbance of methylthiazoletetrazolium by each sample was recorded at 570 nm after staining. (B,C) Cells treated with miR-34a or NC were analyzed by flow cytometry as described in “Materials and methods” for cell cycle distribution analysis.
Mentions: To evaluate the effect of miR-34a in regulating rat liver cell proliferation, a MTT cell proliferation assay and a cell cycle analysis were used (described in “Materials and methods”). Briefly, BRL-3A immortalized rat liver cells were transfected with miR-34a or NC. MTT assay was performed at 2-day intervals, while cell cycle analysis was conducted 48 h after infection. As shown in Figure 2A, miR-34a markedly reduced BRL-3A cell growth at 4 d and more remarkably at 6 d (P<0.01). The growth inhibitory effect of miR-34a can also be sustained by the data of cell cycle analysis, in which the percentages of G2 phase cells in miR-34a and NC groups were (23.14±4.26)% and (8.48±2.93)% respectively, indicating a subpopulation of cells arrested in G2 phase by miR-34a (Figure 2B,C).

Bottom Line: In BRL-3A cells, INHBB was identified as a direct target of miR-34a by luciferase reporter assay.A decrease of INHBB and Met was detected in regenerating liver.MiR-34a expression was upregulated during the late phase of liver regeneration.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Second Military Medical University, Shanghai, China.

ABSTRACT

Background: MicroRNAs are a class of small regulatory RNAs that modulate a variety of biological processes, including cellular differentiation, apoptosis, metabolism and proliferation. This study aims to explore the effect of miR-34a in hepatocyte proliferation and its potential role in liver regeneration termination.

Methodology/principal finding: MiR-34a was highly induced after partial hepatectomy. Overexpression of miR-34a in BRL-3A cells could significantly inhibit cell proliferation and down-regulate the expression of inhibin βB (INHBB) and Met. In BRL-3A cells, INHBB was identified as a direct target of miR-34a by luciferase reporter assay. More importantly, INHBB siRNA significantly repressed cell proliferation. A decrease of INHBB and Met was detected in regenerating liver.

Conclusion/significance: MiR-34a expression was upregulated during the late phase of liver regeneration. MiR-34a-mediated regulation of INHBB and Met may collectively contribute to the suppression of hepatocyte proliferation.

Show MeSH
Related in: MedlinePlus