Open Access

Regulation of glioma migration and invasion via modification of Rap2a activity by the ubiquitin ligase Nedd4-1

  • Authors:
    • Lei Wang
    • Bingxin Zhu
    • Shiquan Wang
    • Yuxuan Wu
    • Wenjian Zhan
    • Shao Xie
    • Hengliang Shi
    • Rutong Yu
  • View Affiliations

  • Published online on: April 11, 2017     https://doi.org/10.3892/or.2017.5572
  • Pages: 2565-2574
  • Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

Νeuronal precursor cell expressed and developmentally downregulated protein (Nedd4-1) is an E3 ubiquitin ligase with critical roles in the pathogenesis of cancer. Herein, we demonstrated that Nedd4-1 protein was upregulated in glioma tissues vs. that in non-cancerous tissues by western blotting and immunohistochemistry. Scratch migration and Transwell chamber assays indicated that downregulation of Nedd4-1 significantly reduced the migration and invasion of the glioma cell lines U251 and U87. Conversely, overexpression of Nedd4-1 obviously enhanced the migratory and invasive capacities in both cell lines. To investigate the role of Nedd4-1 and the intracellular pathways involved, we performed pull-down and co-immunoprecipitation assays, and recognized that Nedd4-1, TNIK and Rap2a formed a complex. Moreover, Nedd4-1 selectively ubiquitinated its specific substrates, the wild-type Rap2a (WT-Rap2a) and dominant-active Rap2a (DA-Rap2a) rather than the dominant-negative Rap2a (DN-Rap2a) in the U251 cells. Subsequently, we demonstrated that Rap2a was robustly ubiquitinated by Nedd4-1 along with the K63-linked, but not the K48-linked ubiquitin chain, which significantly inhibited GTP-Rap2a activity by GST-RalGDS pull-down assay. To further verify whether the ubiquitination of Rap2a by Nedd4-1 regulated the migration and invasion of glioma cells, Nedd4-1, HA-tagged ubiquitin and its mutants as well as WT-Rap2a were co-transfected in the U251 and U87 cell lines. The results confirmed that Nedd4-1 inhibited GTP-Rap2a activity, and promoted the migration and invasion of glioma cells. In brief, our findings demonstrated the important role of Nedd4-1 in regulating the migration and invasion of glioma cells via the Nedd4-1/Rap2a pathway, which may qualify Nedd4-1 as a viable therapeutic target for glioma.
View Figures
View References

Related Articles

Journal Cover

May-2017
Volume 37 Issue 5

Print ISSN: 1021-335X
Online ISSN:1791-2431

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
Spandidos Publications style
Wang L, Zhu B, Wang S, Wu Y, Zhan W, Xie S, Shi H and Yu R: Regulation of glioma migration and invasion via modification of Rap2a activity by the ubiquitin ligase Nedd4-1. Oncol Rep 37: 2565-2574, 2017
APA
Wang, L., Zhu, B., Wang, S., Wu, Y., Zhan, W., Xie, S. ... Yu, R. (2017). Regulation of glioma migration and invasion via modification of Rap2a activity by the ubiquitin ligase Nedd4-1. Oncology Reports, 37, 2565-2574. https://doi.org/10.3892/or.2017.5572
MLA
Wang, L., Zhu, B., Wang, S., Wu, Y., Zhan, W., Xie, S., Shi, H., Yu, R."Regulation of glioma migration and invasion via modification of Rap2a activity by the ubiquitin ligase Nedd4-1". Oncology Reports 37.5 (2017): 2565-2574.
Chicago
Wang, L., Zhu, B., Wang, S., Wu, Y., Zhan, W., Xie, S., Shi, H., Yu, R."Regulation of glioma migration and invasion via modification of Rap2a activity by the ubiquitin ligase Nedd4-1". Oncology Reports 37, no. 5 (2017): 2565-2574. https://doi.org/10.3892/or.2017.5572