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MiRNA-329 targeting E2F1 inhibits cell proliferation in glioma cells.

Xiao B, Tan L, He B, Liu Z, Xu R - J Transl Med (2013)

Bottom Line: The E2F1 was identified as the target of miR-329.Overexpression of miR-329 blocked G1/S transition in LN18 and T98G cell lines, dramatically suppressed cell proliferation and the ability of colony formation.MiR-329 significantly decreased the phosphorylation levels of intracellular kinases Akt and expression of cyclin D1, but the expression of p21 was upregulated, cell growth was suppressed by inhibiting E2F1-mediated Akt pathway.

View Article: PubMed Central - HTML - PubMed

Affiliation: The Neurosurgery Department, General Hospital of Beijing Military Command of People's Liberation Army-PLA, Beijing 100700, P.R. China.

ABSTRACT

Background: MicroRNAs have recently emerged as key regulators of cancers, miR-329 located on 14q32.31 is one of down-regulated miRNAs in glioma, but the function and molecular mechanisms of miR-329 in determining the malignant phenotype of human glioma are elusive. This study therefore was conducted to investigate the role of miR-329 in biological behaviors of human glioma LN18 and T98G cell lines and its molecular mechanisms.

Methods: Nine patients with GBM were analyzed for the expression of miR-329 by quantitative RT-PCR. MiR-329 overexpression was established by transfecting miR-329 precursor into LN18 and T98G cells, and its effects on cell proliferation were studied using MTT assay, anchorage-independent growth ability assay, colony formation assays, Bromodeoxyuridine labeling and immunofluorescence.The effects of miR-329 on cell cycle were studied by flow cytometry. The target of miR-329 was determined by luciferase assays. The regulation of miR-329 on Akt pathway was determined by western blot.

Results: The E2F1 was identified as the target of miR-329. Overexpression of miR-329 blocked G1/S transition in LN18 and T98G cell lines, dramatically suppressed cell proliferation and the ability of colony formation. MiR-329 significantly decreased the phosphorylation levels of intracellular kinases Akt and expression of cyclin D1, but the expression of p21 was upregulated, cell growth was suppressed by inhibiting E2F1-mediated Akt pathway.

Conclusions: MiR-329 may inhibit cell proliferation in human glioma cells through regulating E2F1-mediated suppression of Akt pathway.

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Related in: MedlinePlus

E2F1 mediates suppression of Akt pathway. A) Western blotting analysis of pAkt and Akt after alternation of E2F1 expression. B) Western blotting analysis of pAkt, Akt, p21, cyclinD1 expression after Akt inhibitor IV treatment, β-actin served as the loading control.
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Figure 6: E2F1 mediates suppression of Akt pathway. A) Western blotting analysis of pAkt and Akt after alternation of E2F1 expression. B) Western blotting analysis of pAkt, Akt, p21, cyclinD1 expression after Akt inhibitor IV treatment, β-actin served as the loading control.

Mentions: E2F1 overexpression in glioma cells can cause the phosphorylated level of Akt increase, interfering with the expression of E2F1 can decrease the phosphorylated level of Akt (Figure 6A). The levels of Akt phosphorylation are decreased by treatment with Akt inhibitor IV, in which the p21 is significantly increased and cyclin D1 is downregulated (Figure 6B). These results provided further evidence that miR-329 may negatively regulate the Akt survival pathway through E2F1-mediated suppression of Akt phosphorylation and play an important role in cell proliferation of glioma.


MiRNA-329 targeting E2F1 inhibits cell proliferation in glioma cells.

Xiao B, Tan L, He B, Liu Z, Xu R - J Transl Med (2013)

E2F1 mediates suppression of Akt pathway. A) Western blotting analysis of pAkt and Akt after alternation of E2F1 expression. B) Western blotting analysis of pAkt, Akt, p21, cyclinD1 expression after Akt inhibitor IV treatment, β-actin served as the loading control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750231&req=5

Figure 6: E2F1 mediates suppression of Akt pathway. A) Western blotting analysis of pAkt and Akt after alternation of E2F1 expression. B) Western blotting analysis of pAkt, Akt, p21, cyclinD1 expression after Akt inhibitor IV treatment, β-actin served as the loading control.
Mentions: E2F1 overexpression in glioma cells can cause the phosphorylated level of Akt increase, interfering with the expression of E2F1 can decrease the phosphorylated level of Akt (Figure 6A). The levels of Akt phosphorylation are decreased by treatment with Akt inhibitor IV, in which the p21 is significantly increased and cyclin D1 is downregulated (Figure 6B). These results provided further evidence that miR-329 may negatively regulate the Akt survival pathway through E2F1-mediated suppression of Akt phosphorylation and play an important role in cell proliferation of glioma.

Bottom Line: The E2F1 was identified as the target of miR-329.Overexpression of miR-329 blocked G1/S transition in LN18 and T98G cell lines, dramatically suppressed cell proliferation and the ability of colony formation.MiR-329 significantly decreased the phosphorylation levels of intracellular kinases Akt and expression of cyclin D1, but the expression of p21 was upregulated, cell growth was suppressed by inhibiting E2F1-mediated Akt pathway.

View Article: PubMed Central - HTML - PubMed

Affiliation: The Neurosurgery Department, General Hospital of Beijing Military Command of People's Liberation Army-PLA, Beijing 100700, P.R. China.

ABSTRACT

Background: MicroRNAs have recently emerged as key regulators of cancers, miR-329 located on 14q32.31 is one of down-regulated miRNAs in glioma, but the function and molecular mechanisms of miR-329 in determining the malignant phenotype of human glioma are elusive. This study therefore was conducted to investigate the role of miR-329 in biological behaviors of human glioma LN18 and T98G cell lines and its molecular mechanisms.

Methods: Nine patients with GBM were analyzed for the expression of miR-329 by quantitative RT-PCR. MiR-329 overexpression was established by transfecting miR-329 precursor into LN18 and T98G cells, and its effects on cell proliferation were studied using MTT assay, anchorage-independent growth ability assay, colony formation assays, Bromodeoxyuridine labeling and immunofluorescence.The effects of miR-329 on cell cycle were studied by flow cytometry. The target of miR-329 was determined by luciferase assays. The regulation of miR-329 on Akt pathway was determined by western blot.

Results: The E2F1 was identified as the target of miR-329. Overexpression of miR-329 blocked G1/S transition in LN18 and T98G cell lines, dramatically suppressed cell proliferation and the ability of colony formation. MiR-329 significantly decreased the phosphorylation levels of intracellular kinases Akt and expression of cyclin D1, but the expression of p21 was upregulated, cell growth was suppressed by inhibiting E2F1-mediated Akt pathway.

Conclusions: MiR-329 may inhibit cell proliferation in human glioma cells through regulating E2F1-mediated suppression of Akt pathway.

Show MeSH
Related in: MedlinePlus