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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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Related in: MedlinePlus

Girdin silencing inhibits the expression and activity of MMP-2 and MMP-9. (A and B) Changes in the mRNA levels of MMP-2 and MMP-9 were measured using reverse transcription-quantitative polymerase chain reaction following transfection. The relative mRNA expression levels were calculated using the 2−ΔΔCt method. (C and D) Following transfection, changes in the protein levels of MMP-2 and MMP-9 were detected using western blot analysis. (E and F) Following transfection, gelatin zymography was performed to detect changes in the activities of MMP-2 and MMP-9. Each experiment was repeated three times. The experimental results are presented as the mean ± standard deviation. **P<0.01, compared with the NC group. shRNA, short hairpin RNA; NC, negative control.
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f3-mmr-12-04-5086: Girdin silencing inhibits the expression and activity of MMP-2 and MMP-9. (A and B) Changes in the mRNA levels of MMP-2 and MMP-9 were measured using reverse transcription-quantitative polymerase chain reaction following transfection. The relative mRNA expression levels were calculated using the 2−ΔΔCt method. (C and D) Following transfection, changes in the protein levels of MMP-2 and MMP-9 were detected using western blot analysis. (E and F) Following transfection, gelatin zymography was performed to detect changes in the activities of MMP-2 and MMP-9. Each experiment was repeated three times. The experimental results are presented as the mean ± standard deviation. **P<0.01, compared with the NC group. shRNA, short hairpin RNA; NC, negative control.

Mentions: MMP-2 and MMP-9 are closely associated with cell migration and invasion. To further investigate the effect of girdin silencing on glioma cell proliferation, migration and invasion, RT-qPCR and western blot analyses were performed to detect changes in the expression levels of MMP-2 and MMP-9 following transfection with girdin shRNA. The RT-qPCR results demonstrated that the mRNA levels of MMP-2 and MMP-9 were reduced to 69±8 and 61±8%, respectively, following transfection with girdin shRNA (Fig. 3A and B). Similarly, western blot analysis revealed that the protein expression levels of MMP-2 and MMP-9 were decreased to 65±7 and 58±8% following transfection with girdin shRNA (Fig. 3C and D), which was consistent with the results of the RT-qPCR. As the activities of MMP-2 and MMP-9 are important for their biological functions, gelatin zymography was used to characterize the activities of MMP-2 and MMP-9. The results of the gelatin zymogram revealed that the activities of MMP-2 and MMP-9 were significantly reduced following transfection with girdin shRNA (P<0.01; Fig. 3E and F). These results indicated that girdin silencing inhibited the expression levels and activities of MMP-2 and MMP-9.


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 silencing inhibits the expression and activity of MMP-2 and MMP-9. (A and B) Changes in the mRNA levels of MMP-2 and MMP-9 were measured using reverse transcription-quantitative polymerase chain reaction following transfection. The relative mRNA expression levels were calculated using the 2−ΔΔCt method. (C and D) Following transfection, changes in the protein levels of MMP-2 and MMP-9 were detected using western blot analysis. (E and F) Following transfection, gelatin zymography was performed to detect changes in the activities of MMP-2 and MMP-9. Each experiment was repeated three times. The experimental results are presented as the mean ± standard deviation. **P<0.01, compared with the NC group. shRNA, short hairpin RNA; NC, negative control.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f3-mmr-12-04-5086: Girdin silencing inhibits the expression and activity of MMP-2 and MMP-9. (A and B) Changes in the mRNA levels of MMP-2 and MMP-9 were measured using reverse transcription-quantitative polymerase chain reaction following transfection. The relative mRNA expression levels were calculated using the 2−ΔΔCt method. (C and D) Following transfection, changes in the protein levels of MMP-2 and MMP-9 were detected using western blot analysis. (E and F) Following transfection, gelatin zymography was performed to detect changes in the activities of MMP-2 and MMP-9. Each experiment was repeated three times. The experimental results are presented as the mean ± standard deviation. **P<0.01, compared with the NC group. shRNA, short hairpin RNA; NC, negative control.
Mentions: MMP-2 and MMP-9 are closely associated with cell migration and invasion. To further investigate the effect of girdin silencing on glioma cell proliferation, migration and invasion, RT-qPCR and western blot analyses were performed to detect changes in the expression levels of MMP-2 and MMP-9 following transfection with girdin shRNA. The RT-qPCR results demonstrated that the mRNA levels of MMP-2 and MMP-9 were reduced to 69±8 and 61±8%, respectively, following transfection with girdin shRNA (Fig. 3A and B). Similarly, western blot analysis revealed that the protein expression levels of MMP-2 and MMP-9 were decreased to 65±7 and 58±8% following transfection with girdin shRNA (Fig. 3C and D), which was consistent with the results of the RT-qPCR. As the activities of MMP-2 and MMP-9 are important for their biological functions, gelatin zymography was used to characterize the activities of MMP-2 and MMP-9. The results of the gelatin zymogram revealed that the activities of MMP-2 and MMP-9 were significantly reduced following transfection with girdin shRNA (P<0.01; Fig. 3E and F). These results indicated that girdin silencing inhibited the expression levels and activities of MMP-2 and MMP-9.

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