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MiR-152 suppresses the proliferation and invasion of NSCLC cells by inhibiting FGF2.

Cheng Z, Ma R, Tan W, Zhang L - Exp. Mol. Med. (2014)

Bottom Line: MicroRNAs (miRNAs) regulate the proliferation and metastasis of cancer cells.Overexpression of miR-152 suppressed cell proliferation and colony formation and also limited migration and invasion.FGF2 knockdown suppressed cell proliferation, colony formation, migration and invasion, whereas FGF2 overexpression partially reversed the suppressive effect of miR-152.

View Article: PubMed Central - PubMed

Affiliation: Department of Respiratory Medicine, ZhongNan Hospital of WuHan University, WuHan, China.

ABSTRACT
MicroRNAs (miRNAs) regulate the proliferation and metastasis of cancer cells. Here, we showed that miR-152 was downregulated in non-small-cell lung cancer (NSCLC) tissues and cell lines. Overexpression of miR-152 suppressed cell proliferation and colony formation and also limited migration and invasion. Fibroblast growth factor 2 (FGF2) was confirmed as a direct target of miR-152. FGF2 knockdown suppressed cell proliferation, colony formation, migration and invasion, whereas FGF2 overexpression partially reversed the suppressive effect of miR-152. Furthermore, the presence of miR-152 was inversely correlated with FGF2 in NSCLC tissues. Overall, this study demonstrated that miR-152 suppressed the proliferation and invasion of NSCLC cells by downregulating FGF2. These findings provide novel insights with potential therapeutic applications for the treatment of NSCLC.

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Fibroblast growth factor 2 (FGF2) was a direct target of microRNA-152 (miR-152). (a) Schematic representation of wild-type (WT) and mutated (Mut) putative miR-152-binding sites in the 3′-untranslated region (3′-UTR) of FGF2. (b) HEK293 cells were co-transfected with 100 nM WT or Mut FGF2 3'-UTR, pGL-3 control and either miR-152 or negative control mimics. Relative firefly luciferase activity was normalized to Renilla luciferase activity. (c) Expression of FGF2 mRNA was detected by quantitative real-time PCR (qRT-PCR) in A549 cells transfected with miR-152 or control mimic and in A549 cells transfected miR-152 or control inhibitor. (d) Protein levels were detected by western blot analysis in A549 cells transfected with miR-152 or control mimic and in A549 cells transfected with miR-152 or control inhibitor. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal control. *P<0.05, **P<0.01 vs control.
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fig4: Fibroblast growth factor 2 (FGF2) was a direct target of microRNA-152 (miR-152). (a) Schematic representation of wild-type (WT) and mutated (Mut) putative miR-152-binding sites in the 3′-untranslated region (3′-UTR) of FGF2. (b) HEK293 cells were co-transfected with 100 nM WT or Mut FGF2 3'-UTR, pGL-3 control and either miR-152 or negative control mimics. Relative firefly luciferase activity was normalized to Renilla luciferase activity. (c) Expression of FGF2 mRNA was detected by quantitative real-time PCR (qRT-PCR) in A549 cells transfected with miR-152 or control mimic and in A549 cells transfected miR-152 or control inhibitor. (d) Protein levels were detected by western blot analysis in A549 cells transfected with miR-152 or control mimic and in A549 cells transfected with miR-152 or control inhibitor. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal control. *P<0.05, **P<0.01 vs control.

Mentions: To identify targets of miR-152, we used TargetScan 6.2 (http://www.targetscan.org), a widely used miRNA target prediction website. FGF2 was found to be a potential target (Figure 4a). Luciferase activity assays are commonly used to validate the suppressive effects of miRNAs on their target mRNAs. Here, we found that miR-152 significantly inhibited the luciferase activity of the wild-type but not the Mut 3′-UTR of FGF2 in HEK293 cells (Figure 4b). Moreover, overexpression of miR-152 significantly suppressed levels of both FGF2 mRNA and protein, whereas the inhibition of miR-152 significantly increased these levels (Figures 4c and d). These data suggest that miR-152 might induce the degradation of FGF2 mRNA, leading to a reduction in levels of FGF2 protein and implying that FGF2 is a direct target of miR-152.


MiR-152 suppresses the proliferation and invasion of NSCLC cells by inhibiting FGF2.

Cheng Z, Ma R, Tan W, Zhang L - Exp. Mol. Med. (2014)

Fibroblast growth factor 2 (FGF2) was a direct target of microRNA-152 (miR-152). (a) Schematic representation of wild-type (WT) and mutated (Mut) putative miR-152-binding sites in the 3′-untranslated region (3′-UTR) of FGF2. (b) HEK293 cells were co-transfected with 100 nM WT or Mut FGF2 3'-UTR, pGL-3 control and either miR-152 or negative control mimics. Relative firefly luciferase activity was normalized to Renilla luciferase activity. (c) Expression of FGF2 mRNA was detected by quantitative real-time PCR (qRT-PCR) in A549 cells transfected with miR-152 or control mimic and in A549 cells transfected miR-152 or control inhibitor. (d) Protein levels were detected by western blot analysis in A549 cells transfected with miR-152 or control mimic and in A549 cells transfected with miR-152 or control inhibitor. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal control. *P<0.05, **P<0.01 vs control.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4150934&req=5

fig4: Fibroblast growth factor 2 (FGF2) was a direct target of microRNA-152 (miR-152). (a) Schematic representation of wild-type (WT) and mutated (Mut) putative miR-152-binding sites in the 3′-untranslated region (3′-UTR) of FGF2. (b) HEK293 cells were co-transfected with 100 nM WT or Mut FGF2 3'-UTR, pGL-3 control and either miR-152 or negative control mimics. Relative firefly luciferase activity was normalized to Renilla luciferase activity. (c) Expression of FGF2 mRNA was detected by quantitative real-time PCR (qRT-PCR) in A549 cells transfected with miR-152 or control mimic and in A549 cells transfected miR-152 or control inhibitor. (d) Protein levels were detected by western blot analysis in A549 cells transfected with miR-152 or control mimic and in A549 cells transfected with miR-152 or control inhibitor. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal control. *P<0.05, **P<0.01 vs control.
Mentions: To identify targets of miR-152, we used TargetScan 6.2 (http://www.targetscan.org), a widely used miRNA target prediction website. FGF2 was found to be a potential target (Figure 4a). Luciferase activity assays are commonly used to validate the suppressive effects of miRNAs on their target mRNAs. Here, we found that miR-152 significantly inhibited the luciferase activity of the wild-type but not the Mut 3′-UTR of FGF2 in HEK293 cells (Figure 4b). Moreover, overexpression of miR-152 significantly suppressed levels of both FGF2 mRNA and protein, whereas the inhibition of miR-152 significantly increased these levels (Figures 4c and d). These data suggest that miR-152 might induce the degradation of FGF2 mRNA, leading to a reduction in levels of FGF2 protein and implying that FGF2 is a direct target of miR-152.

Bottom Line: MicroRNAs (miRNAs) regulate the proliferation and metastasis of cancer cells.Overexpression of miR-152 suppressed cell proliferation and colony formation and also limited migration and invasion.FGF2 knockdown suppressed cell proliferation, colony formation, migration and invasion, whereas FGF2 overexpression partially reversed the suppressive effect of miR-152.

View Article: PubMed Central - PubMed

Affiliation: Department of Respiratory Medicine, ZhongNan Hospital of WuHan University, WuHan, China.

ABSTRACT
MicroRNAs (miRNAs) regulate the proliferation and metastasis of cancer cells. Here, we showed that miR-152 was downregulated in non-small-cell lung cancer (NSCLC) tissues and cell lines. Overexpression of miR-152 suppressed cell proliferation and colony formation and also limited migration and invasion. Fibroblast growth factor 2 (FGF2) was confirmed as a direct target of miR-152. FGF2 knockdown suppressed cell proliferation, colony formation, migration and invasion, whereas FGF2 overexpression partially reversed the suppressive effect of miR-152. Furthermore, the presence of miR-152 was inversely correlated with FGF2 in NSCLC tissues. Overall, this study demonstrated that miR-152 suppressed the proliferation and invasion of NSCLC cells by downregulating FGF2. These findings provide novel insights with potential therapeutic applications for the treatment of NSCLC.

Show MeSH
Related in: MedlinePlus