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Girdin regulates the migration and invasion of glioma cells via the PI3K-Akt signaling pathway.

Ni W, Fang Y, Tong L, Tong Z, Yi F, Qiu J, Wang R, Tong X - Mol Med Rep (2015)

Bottom Line: In the present study, short‑hairpin RNA technology was used to silence the gene expression of girdin.In addition, the expression levels and activity of matrix metalloproteinase (MMP)‑2 and MMP‑9 were also affected by girdin silencing.Therefore, the results of the present study provide a theoretical foundation for the development of anticancer drugs.

View Article: PubMed Central - PubMed

Affiliation: Department of Human Anatomy, Histology and Embryology, China Medical University, Shenyang, Liaoning 110001, P.R. China.

ABSTRACT
Girdin, an actin‑binding protein, is associated with cell migration and is expressed at high levels in glioma cells. However, the association between girdin and the development of glioma remains to be elucidated. In the present study, short‑hairpin RNA technology was used to silence the gene expression of girdin. The effects of girdin silencing on glioma cell proliferation, migration and invasion were then assessed using a cell viability assay, wound‑healing assay, transwell invasion assay, reverse transcription‑quantitative polymerase chain reaction, western blot analysis and gelatin zymography. The results suggested that girdin silencing inhibited the proliferation, migration and invasion of glioma cells. In addition, the expression levels and activity of matrix metalloproteinase (MMP)‑2 and MMP‑9 were also affected by girdin silencing. Further mechanistic investigation indicated that girdin may regulate glioma cell migration and invasion through the phosphatidylinositol‑3‑kinase/protein kinase B (PI3K‑Akt) signaling pathway. Therefore, the results of the present study provide a theoretical foundation for the development of anticancer drugs.

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Girdin regulates glioma cell migration and invasion via the PI3K-Akt signaling pathway. (A and B) Western blot analysis was performed to detect changes in the levels of p85α, p110α, p-Akt and Akt following transfection with girdin shRNA. (C and D) Western blot analysis was performed to detect the effects of girdin silencing on the protein levels of p-Akt, Akt and MMP-9 following treatment with LY294002. (E and F) A Transwell invasion assay was performed to detect the effect of girdin silencing on cell invasion following treatment with LY294002. Images were captured under a light microscope (magnification, ×200). Each experiment was repeated three times. The experimental results are presented as the mean ± standard deviation. **P<0.01, compared with the negative control. shRNA, short hairpin RNA; NC, negative control; DMSO, dimethyl sulfoxide; MMP, matrix metalloproteinase; Akt, protein kinase B; p-, phosphorylated.
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f4-mmr-12-04-5086: Girdin regulates glioma cell migration and invasion via the PI3K-Akt signaling pathway. (A and B) Western blot analysis was performed to detect changes in the levels of p85α, p110α, p-Akt and Akt following transfection with girdin shRNA. (C and D) Western blot analysis was performed to detect the effects of girdin silencing on the protein levels of p-Akt, Akt and MMP-9 following treatment with LY294002. (E and F) A Transwell invasion assay was performed to detect the effect of girdin silencing on cell invasion following treatment with LY294002. Images were captured under a light microscope (magnification, ×200). Each experiment was repeated three times. The experimental results are presented as the mean ± standard deviation. **P<0.01, compared with the negative control. shRNA, short hairpin RNA; NC, negative control; DMSO, dimethyl sulfoxide; MMP, matrix metalloproteinase; Akt, protein kinase B; p-, phosphorylated.

Mentions: To further investigate how girdin affects glioma cell migration and invasion, western blotting was used to detect the protein levels of p85α, p110α, Akt and p-Akt following transfection with girdin shRNA. The results demonstrated that the protein levels of p85α, p110α and p-Akt were reduced to 55±9, 58±7 and 59±8%, respectively; however, no significant changes were detected in the expression of Akt (Fig. 4A and B). These results indicated that girdin silencing affected the activation of the PI3K-Akt signaling pathway. The LY294002 PI3K signaling pathway inhibitor was used to inhibit the PI3K-Akt pathway, followed by western blot analysis to detect changes in the levels of Akt, p-Akt and MMP-9. The results revealed that the levels of p-Akt and MMP-9 were significantly reduced following girdin silencing. In addition, the LY294002-mediated PI3K-Akt signaling pathway inhibition yielded the same results as girdin silencing. The combined application of LY294002 with girdin silencing further enhanced the reductions of p-Akt and MMP-9 (Fig. 4C and D). A Transwell invasion assay, which was used to detect the effect of LY294002 on glioma cell invasion, also demonstrated that LY294002 further enhanced the inhibitory effect of girdin silencing on glioma cell invasion (Fig. 4E and F). These results suggested that girdin silencing may affect glioma cell migration and invasion by regulating the PI3K-Akt signaling pathway.


Girdin regulates the migration and invasion of glioma cells via the PI3K-Akt signaling pathway.

Ni W, Fang Y, Tong L, Tong Z, Yi F, Qiu J, Wang R, Tong X - Mol Med Rep (2015)

Girdin regulates glioma cell migration and invasion via the PI3K-Akt signaling pathway. (A and B) Western blot analysis was performed to detect changes in the levels of p85α, p110α, p-Akt and Akt following transfection with girdin shRNA. (C and D) Western blot analysis was performed to detect the effects of girdin silencing on the protein levels of p-Akt, Akt and MMP-9 following treatment with LY294002. (E and F) A Transwell invasion assay was performed to detect the effect of girdin silencing on cell invasion following treatment with LY294002. Images were captured under a light microscope (magnification, ×200). Each experiment was repeated three times. The experimental results are presented as the mean ± standard deviation. **P<0.01, compared with the negative control. shRNA, short hairpin RNA; NC, negative control; DMSO, dimethyl sulfoxide; MMP, matrix metalloproteinase; Akt, protein kinase B; p-, phosphorylated.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581799&req=5

f4-mmr-12-04-5086: Girdin regulates glioma cell migration and invasion via the PI3K-Akt signaling pathway. (A and B) Western blot analysis was performed to detect changes in the levels of p85α, p110α, p-Akt and Akt following transfection with girdin shRNA. (C and D) Western blot analysis was performed to detect the effects of girdin silencing on the protein levels of p-Akt, Akt and MMP-9 following treatment with LY294002. (E and F) A Transwell invasion assay was performed to detect the effect of girdin silencing on cell invasion following treatment with LY294002. Images were captured under a light microscope (magnification, ×200). Each experiment was repeated three times. The experimental results are presented as the mean ± standard deviation. **P<0.01, compared with the negative control. shRNA, short hairpin RNA; NC, negative control; DMSO, dimethyl sulfoxide; MMP, matrix metalloproteinase; Akt, protein kinase B; p-, phosphorylated.
Mentions: To further investigate how girdin affects glioma cell migration and invasion, western blotting was used to detect the protein levels of p85α, p110α, Akt and p-Akt following transfection with girdin shRNA. The results demonstrated that the protein levels of p85α, p110α and p-Akt were reduced to 55±9, 58±7 and 59±8%, respectively; however, no significant changes were detected in the expression of Akt (Fig. 4A and B). These results indicated that girdin silencing affected the activation of the PI3K-Akt signaling pathway. The LY294002 PI3K signaling pathway inhibitor was used to inhibit the PI3K-Akt pathway, followed by western blot analysis to detect changes in the levels of Akt, p-Akt and MMP-9. The results revealed that the levels of p-Akt and MMP-9 were significantly reduced following girdin silencing. In addition, the LY294002-mediated PI3K-Akt signaling pathway inhibition yielded the same results as girdin silencing. The combined application of LY294002 with girdin silencing further enhanced the reductions of p-Akt and MMP-9 (Fig. 4C and D). A Transwell invasion assay, which was used to detect the effect of LY294002 on glioma cell invasion, also demonstrated that LY294002 further enhanced the inhibitory effect of girdin silencing on glioma cell invasion (Fig. 4E and F). These results suggested that girdin silencing may affect glioma cell migration and invasion by regulating the PI3K-Akt signaling pathway.

Bottom Line: In the present study, short‑hairpin RNA technology was used to silence the gene expression of girdin.In addition, the expression levels and activity of matrix metalloproteinase (MMP)‑2 and MMP‑9 were also affected by girdin silencing.Therefore, the results of the present study provide a theoretical foundation for the development of anticancer drugs.

View Article: PubMed Central - PubMed

Affiliation: Department of Human Anatomy, Histology and Embryology, China Medical University, Shenyang, Liaoning 110001, P.R. China.

ABSTRACT
Girdin, an actin‑binding protein, is associated with cell migration and is expressed at high levels in glioma cells. However, the association between girdin and the development of glioma remains to be elucidated. In the present study, short‑hairpin RNA technology was used to silence the gene expression of girdin. The effects of girdin silencing on glioma cell proliferation, migration and invasion were then assessed using a cell viability assay, wound‑healing assay, transwell invasion assay, reverse transcription‑quantitative polymerase chain reaction, western blot analysis and gelatin zymography. The results suggested that girdin silencing inhibited the proliferation, migration and invasion of glioma cells. In addition, the expression levels and activity of matrix metalloproteinase (MMP)‑2 and MMP‑9 were also affected by girdin silencing. Further mechanistic investigation indicated that girdin may regulate glioma cell migration and invasion through the phosphatidylinositol‑3‑kinase/protein kinase B (PI3K‑Akt) signaling pathway. Therefore, the results of the present study provide a theoretical foundation for the development of anticancer drugs.

Show MeSH
Related in: MedlinePlus