Limits...
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.

Show MeSH

Related in: MedlinePlus

Girdin shRNA decreases the gene expression of girdin. (A and B) Protein expression levels of girdin in the U373, U251, A172, U87-MG and SHG-44 cell lines were detected using western blot, with β-actin as a reference. (C) Changes in mRNA levels of girdin were detected using reverse transcription-quantitative polymerase chain reaction, following transfection with girdin shRNA. The relative mRNA expression level in each sample was quantified using the 2−∆∆Ct method. (D and E) Expression levels of girdin were measured using western blot analysis following transfection. Each experiment was repeated three times. The experimental results are presented as the mean ± standard deviation. ***P<0.001, compared with the NC group. shRNA, short hairpin RNA; NC, negative control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4581799&req=5

f1-mmr-12-04-5086: Girdin shRNA decreases the gene expression of girdin. (A and B) Protein expression levels of girdin in the U373, U251, A172, U87-MG and SHG-44 cell lines were detected using western blot, with β-actin as a reference. (C) Changes in mRNA levels of girdin were detected using reverse transcription-quantitative polymerase chain reaction, following transfection with girdin shRNA. The relative mRNA expression level in each sample was quantified using the 2−∆∆Ct method. (D and E) Expression levels of girdin were measured using western blot analysis following transfection. Each experiment was repeated three times. The experimental results are presented as the mean ± standard deviation. ***P<0.001, compared with the NC group. shRNA, short hairpin RNA; NC, negative control.

Mentions: To select the appropriate cell line for the present study, the expression levels of girdin in U373, U251, A172, U87-MG and SHG-44 cell lines were assessed using western blot analysis. As the U251 cell line exhibited the highest expression level of girdin (Fig. 1A and B), the U251 cells were selected for use in the subsequent experiments. Following transfection with girdin shRNA, the expression of girdin was detected using RT-qPCR and western blot analysis. The RT-qPCR results indicated that the mRNA levels of girdin were reduced to 25±4% following transfection with girdin shRNA (Fig. 1C). Similarly, the western blot analysis revealed that the protein expression of girdin was significantly reduced following transfection with girdin shRNA (P<0.001; Fig. 1D and E). These results suggested that the expression of girdin was effectively silenced following transfection with girdin shRNA.


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 shRNA decreases the gene expression of girdin. (A and B) Protein expression levels of girdin in the U373, U251, A172, U87-MG and SHG-44 cell lines were detected using western blot, with β-actin as a reference. (C) Changes in mRNA levels of girdin were detected using reverse transcription-quantitative polymerase chain reaction, following transfection with girdin shRNA. The relative mRNA expression level in each sample was quantified using the 2−∆∆Ct method. (D and E) Expression levels of girdin were measured using western blot analysis following transfection. Each experiment was repeated three times. The experimental results are presented as the mean ± standard deviation. ***P<0.001, 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

f1-mmr-12-04-5086: Girdin shRNA decreases the gene expression of girdin. (A and B) Protein expression levels of girdin in the U373, U251, A172, U87-MG and SHG-44 cell lines were detected using western blot, with β-actin as a reference. (C) Changes in mRNA levels of girdin were detected using reverse transcription-quantitative polymerase chain reaction, following transfection with girdin shRNA. The relative mRNA expression level in each sample was quantified using the 2−∆∆Ct method. (D and E) Expression levels of girdin were measured using western blot analysis following transfection. Each experiment was repeated three times. The experimental results are presented as the mean ± standard deviation. ***P<0.001, compared with the NC group. shRNA, short hairpin RNA; NC, negative control.
Mentions: To select the appropriate cell line for the present study, the expression levels of girdin in U373, U251, A172, U87-MG and SHG-44 cell lines were assessed using western blot analysis. As the U251 cell line exhibited the highest expression level of girdin (Fig. 1A and B), the U251 cells were selected for use in the subsequent experiments. Following transfection with girdin shRNA, the expression of girdin was detected using RT-qPCR and western blot analysis. The RT-qPCR results indicated that the mRNA levels of girdin were reduced to 25±4% following transfection with girdin shRNA (Fig. 1C). Similarly, the western blot analysis revealed that the protein expression of girdin was significantly reduced following transfection with girdin shRNA (P<0.001; Fig. 1D and E). These results suggested that the expression of girdin was effectively silenced following transfection with girdin shRNA.

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