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RhoC GTPase Activation Assay
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The ubiquitin ligase RNF8 regulates Rho GTPases and promotes cytoskeletal changes and motility in triple-negative

Bruno Pereira De Carvalho1, Yi-Jye Chern1, Jiabei He1

  • 1Department of Pharmacological Sciences, Stony Brook University, NY, USA.

FEBS Letters
|November 18, 2020
PubMed

Insights

The ubiquitin ligase RNF8 regulates cell migration in triple-negative breast cancer (TNBC) by controlling Rho GTPases. RNF8 knockdown inhibits metastasis-associated proteins RHOA and CDC42, reducing cell motility.

Area of Science:

  • Oncology
  • Molecular Biology
  • Cell Biology

Background:

  • The ubiquitin ligase RNF8 is implicated in epithelial-to-mesenchymal transition (EMT) and metastasis in triple-negative breast cancer (TNBC).
  • Rho GTPases are critical regulators of cancer cell metastasis.

Purpose of the Study:

  • To investigate the role of RNF8 in regulating Rho GTPases and cell motility in TNBC.
  • To elucidate the molecular mechanisms by which RNF8 influences cancer cell migration.

Main Methods:

  • RNF8 knockdown was performed in TNBC cells.
  • Protein and mRNA levels of Rho GTPases (RHOA, CDC42) were assessed.
  • Cellular structures involved in migration, such as filopodia and focal adhesions, were analyzed.
  • Cell migration assays were conducted.

Main Results:

  • RNF8 knockdown significantly reduced the protein and mRNA levels of RHOA and CDC42 in TNBC cells.
  • The formation of filopodia and focal adhesions, and their linkage to stress fibers, were impaired following RNF8 knockdown.
  • Cell migration was markedly inhibited in RNF8-depleted TNBC cells.

Conclusions:

  • RNF8 plays a crucial role in regulating cell migration in TNBC.
  • RNF8 mediates TNBC cell migration through the modulation of Rho GTPases RHOA and CDC42.
  • RNF8 represents a potential therapeutic target for inhibiting metastasis in TNBC.
Keywords:
CDC42RHOARNF8cell migrationcytoskeleton rearrangementsfilopodiafocal adhesiontriple-negative breast cancer

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