title: Rho GTPases RhoA and Rac1 mediate effects of dietary folate on metastatic potential of A549 cancer cells through the control of cofilin phosphorylation.
abstract: 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.
authors.affiliation: Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, South Carolina 29425.
authors.forename: Natalia V
authors.initials: NV
authors.lastname: Oleinik
authors.affiliation: Comparative Medicine and Laboratory Animal Resources, and Medical University of South Carolina, Charleston, South Carolina 29425.
authors.forename: Kristi L
authors.initials: KL
authors.lastname: Helke
authors.affiliation: Hollings Cancer Center, Medical University of South Carolina, Charleston, South Carolina 29425.
authors.forename: Emily
authors.initials: E
authors.lastname: Kistner-Griffin
authors.affiliation: Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, South Carolina 29425; Hollings Cancer Center, Medical University of South Carolina, Charleston, South Carolina 29425.
authors.forename: Natalia I
authors.initials: NI
authors.lastname: Krupenko
authors.affiliation: Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, South Carolina 29425; Hollings Cancer Center, Medical University of South Carolina, Charleston, South Carolina 29425. Electronic address: sergey_krupenko@unc.edu.
authors.forename: Sergey A
authors.initials: SA
authors.lastname: Krupenko
mesh_terms: D000230:Adenocarcinoma / Q000201:enzymology* / Q000556:secondary; D000284:Administration, Oral; D000818:Animals; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000201:enzymology; D002465:Cell Movement / Q000187:drug effects; D002470:Cell Survival; D051338:Cofilin 1 / Q000378:metabolism*; D019587:Dietary Supplements; D004721:Endoplasmic Reticulum / Q000201:enzymology; D058750:Epithelial-Mesenchymal Transition; D005492:Folic Acid / Q000008:administration & dosage* / Q000494:pharmacology; D006801:Humans; D008175:Lung Neoplasms / Q000201:enzymology* / Q000473:pathology; D008207:Lymphatic Metastasis; D008297:Male; D008745:Methylation; D016513:Mice, SCID; D009368:Neoplasm Transplantation; D010766:Phosphorylation; D054730:Protein Interaction Domains and Motifs; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D015398:Signal Transduction; D020764:cdc42 GTP-Binding Protein / Q000378:metabolism; D054462:p21-Activated Kinases / Q000737:chemistry; D020830:rac1 GTP-Binding Protein / Q000737:chemistry / Q000502:physiology*; D020742:rhoA GTP-Binding Protein / Q000502:physiology*
chemical_list: C495810:CFL1 protein, human; D051338:Cofilin 1; C107953:RAC1 protein, human; C066737:RHOA protein, human; D005492:Folic Acid; D054462:p21-Activated Kinases; D020764:cdc42 GTP-Binding Protein; D020830:rac1 GTP-Binding Protein; D020742:rhoA GTP-Binding Protein
keywords: Cell Motility; Cofilin; Folate; Rho GTPases; Tumor Metastases
references.citation: Cancer Res. 2014 Apr 15;74(8):2362-73
references.pmid: 24509905
references.citation: Am J Physiol Cell Physiol. 2009 Mar;296(3):C463-75
references.pmid: 19109524
references.citation: Science. 2003 Dec 5;302(5651):1704-9
references.pmid: 14657486
references.citation: J Cell Biochem. 2009 Dec 15;108(6):1252-62
references.pmid: 19862699
references.citation: Hum Mol Genet. 2010 Apr 1;19(7):1347-57
references.pmid: 20067919
references.citation: Int J Cancer. 1993 Apr 22;54(1):112-8
references.pmid: 8478137
references.citation: Oncogene. 2010 Nov 25;29(47):6233-44
references.pmid: 20729910
references.citation: EMBO J. 2006 Feb 22;25(4):713-26
references.pmid: 16456544
references.citation: J Biol Chem. 2005 Mar 25;280(12):11448-57
references.pmid: 15647284
references.citation: PLoS One. 2013 Jul 24;8(7):e69389
references.pmid: 23894465
references.citation: Arch Biochem Biophys. 1991 May 1;286(2):633-7
references.pmid: 1897982
references.citation: Nat Rev Mol Cell Biol. 2008 Sep;9(9):690-701
references.pmid: 18719708
references.citation: Carcinogenesis. 2012 Jun;33(6):1158-68
references.pmid: 22461522
references.citation: Cancer Epidemiol. 2012 Feb;36(1):78-81
references.pmid: 22018948
references.citation: J Nutr. 2009 Dec;139(12):2402-5
references.pmid: 19812215
references.citation: Cancer Lett. 2002 Dec 10;187(1-2):69-75
references.pmid: 12359353
references.citation: Free Radic Biol Med. 2003 Aug 15;35(4):428-37
references.pmid: 12899944
references.citation: Cell Adh Migr. 2011 Jan-Feb;5(1):11-5
references.pmid: 20798596
references.citation: Curr Opin Clin Nutr Metab Care. 2009 Nov;12(6):555-64
references.pmid: 19726978
references.citation: JAMA. 2007 Jun 6;297(21):2408-9
references.pmid: 17551134
references.citation: Vitam Horm. 2008;79:1-44
references.pmid: 18804690
references.citation: Nat Rev Immunol. 2009 Sep;9(9):630-44
references.pmid: 19696767
references.citation: J Clin Invest. 2009 Jun;119(6):1420-8
references.pmid: 19487818
references.citation: Biochem J. 2000 Jun 1;348 Pt 2:241-55
references.pmid: 10816416
references.citation: Nat Rev Cancer. 2003 May;3(5):362-74
references.pmid: 12724734
references.citation: Cancer Metastasis Rev. 2012 Dec;31(3-4):441-54
references.pmid: 22692480
references.citation: J Biol Chem. 2009 Oct 9;284(41):27964-27973
references.pmid: 19651782
references.citation: Genes Cancer. 2011 Sep;2(9):889-99
references.pmid: 22593801
references.citation: Nature. 2002 Dec 12;420(6916):629-35
references.pmid: 12478284
references.citation: Nat Rev Cancer. 2006 May;6(5):392-401
references.pmid: 16572188
references.citation: Exp Oncol. 2010 Sep;32(3):137-42
references.pmid: 21403607
references.citation: Biochem Pharmacol. 2006 Jul 14;72(2):256-66
references.pmid: 16712799
references.citation: Respir Res. 2005 Jun 09;6:56
references.pmid: 15946381
references.citation: Nat Rev Cancer. 2003 Jun;3(6):453-8
references.pmid: 12778135
references.citation: J Mol Med (Berl). 2007 Jun;85(6):555-68
references.pmid: 17294230
references.citation: Br J Pharmacol. 2012 Aug;166(7):2036-48
references.pmid: 22300324
references.citation: Mol Cell. 2013 Feb 7;49(3):379-87
references.pmid: 23395268
references.citation: J Biol Chem. 2001 Feb 23;276(8):5841-5
references.pmid: 11121396
references.citation: Cell. 2011 Oct 14;147(2):275-92
references.pmid: 22000009
references.citation: Nature. 1998 Jun 25;393(6687):809-12
references.pmid: 9655398
references.citation: Curr Opin Cell Biol. 2006 Feb;18(1):26-31
references.pmid: 16337782
references.citation: Cancer Discov. 2013 Jun;3(6):616-29
references.pmid: 23585416
references.citation: Proc Natl Acad Sci U S A. 2003 May 27;100(11):6529-34
references.pmid: 12750467
references.citation: J Biol Chem. 2000 Jun 9;275(23):17605-10
references.pmid: 10747846
references.citation: PLoS One. 2013 May 28;8(5):e65189
references.pmid: 23724131
references.citation: Biochem Soc Symp. 2006;(73):85-96
references.pmid: 16626290
references.citation: Lancet. 2013 Mar 23;381(9871):1029-36
references.pmid: 23352552
references.citation: Curr Opin Investig Drugs. 2010 Dec;11(12):1409-23
references.pmid: 21154123
references.citation: J Histochem Cytochem. 2003 Apr;51(4):411-20
references.pmid: 12642619
references.citation: JAMA. 2009 Nov 18;302(19):2119-26
references.pmid: 19920236
references.citation: Clin Chem Lab Med. 2013 Mar 1;51(3):607-16
references.pmid: 23241610
references.citation: Am J Clin Nutr. 1995 Jan;61(1):90-6
references.pmid: 7529961
references.citation: Biochem Pharmacol. 2013 Feb 1;85(3):376-84
references.pmid: 23178654
references.citation: Cell. 2007 Jun 1;129(5):865-77
references.pmid: 17540168
references.citation: Nat Protoc. 2009;4(11):1623-31
references.pmid: 19834477
references.citation: Trends Pharmacol Sci. 2001 Jan;22(1):32-9
references.pmid: 11165670
references.citation: J Neurochem. 2012 Jun;121(6):903-14
references.pmid: 22428561
references.citation: EMBO J. 2004 Dec 8;23(24):4792-801
references.pmid: 15538389
references.citation: J Biol Chem. 2010 Jul 23;285(30):23056-63
references.pmid: 20498374
references.citation: Biochem Pharmacol. 1994 Jul 19;48(2):319-25
references.pmid: 8053927
references.citation: PLoS One. 2012;7(11):e47201
references.pmid: 23144806
references.citation: Nat Rev Cancer. 2005 May;5(5):405-12
references.pmid: 15864282
references.citation: Mol Pharmacol. 2001 Dec;60(6):1288-95
references.pmid: 11723236
references.citation: Subcell Biochem. 2012;56:105-30
references.pmid: 22116697
references.citation: Nat Rev Cancer. 2007 Jun;7(6):429-40
references.pmid: 17522712
references.citation: Am J Clin Nutr. 2008 Mar;87(3):517-33
references.pmid: 18326588