Rho GTPases RhoA and Rac1 mediate effects of dietary folate on metastatic potential of A549 cancer cells through the control of cofilin phosphorylation

J Biol Chem. 2014 Sep 19;289(38):26383-26394. doi: 10.1074/jbc.M114.569657. Epub 2014 Aug 1.


Folate, an important nutrient in the human diet, has been implicated in cancer, but its role in metastasis is not established. We have shown previously that the withdrawal of medium folate leads to the inhibition of migration and invasion of A549 lung carcinoma cells. Here we have demonstrated that medium folate regulates the function of Rho GTPases by enabling their carboxyl methylation and translocation to plasma membrane. Conversely, the lack of folate leads to the retention of these proteins in endoplasmic reticulum. Folate also promoted the switch from inactive (GDP-bound) to active (GTP-bound) GTPases, resulting in the activation of downstream kinases p21-activated kinase and LIM kinase and phosphorylation of the actin-depolymerizing factor cofilin. We have further demonstrated that in A549 cells two GTPases, RhoA and Rac1, but not Cdc42, are immediate sensors of folate status: the siRNA silencing of RhoA or Rac1 blocked effects of folate on cofilin phosphorylation and cellular migration and invasion. The finding that folate modulates metastatic potential of cancer cells was confirmed in an animal model of lung cancer using tail vein injection of A549 cells in SCID mice. A folate-rich diet enhanced lung colonization and distant metastasis to lymph nodes and decreased overall survival (35 versus 63 days for mice on a folate-restricted diet). High folate also promoted epithelial-mesenchymal transition in cancer cells and experimental mouse tumors. Our study provides experimental evidence for a mechanism of metastasis promotion by dietary folate and highlights the interaction between nutrients and metastasis-related signaling.

Keywords: Cell Motility; Cofilin; Folate; Rho GTPases; Tumor Metastases.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adenocarcinoma / enzymology*
  • Adenocarcinoma / secondary
  • Administration, Oral
  • Animals
  • Cell Line, Tumor
  • Cell Membrane / enzymology
  • Cell Movement / drug effects
  • Cell Survival
  • Cofilin 1 / metabolism*
  • Dietary Supplements
  • Endoplasmic Reticulum / enzymology
  • Epithelial-Mesenchymal Transition
  • Folic Acid / administration & dosage*
  • Folic Acid / pharmacology
  • Humans
  • Lung Neoplasms / enzymology*
  • Lung Neoplasms / pathology
  • Lymphatic Metastasis
  • Male
  • Methylation
  • Mice, SCID
  • Neoplasm Transplantation
  • Phosphorylation
  • Protein Interaction Domains and Motifs
  • Protein Processing, Post-Translational
  • Protein Transport
  • Signal Transduction
  • cdc42 GTP-Binding Protein / metabolism
  • p21-Activated Kinases / chemistry
  • rac1 GTP-Binding Protein / chemistry
  • rac1 GTP-Binding Protein / physiology*
  • rhoA GTP-Binding Protein / physiology*


  • CFL1 protein, human
  • Cofilin 1
  • RAC1 protein, human
  • RHOA protein, human
  • Folic Acid
  • p21-Activated Kinases
  • cdc42 GTP-Binding Protein
  • rac1 GTP-Binding Protein
  • rhoA GTP-Binding Protein