title: Effect of down-regulation of aquaporins in human corneal endothelial and epithelial cell lines.
abstract: OBJECTIVE
  The purpose of this study was to determine the effects of down-regulation of Aquaporin 1 (AQP1) and Aquaporin 5 (AQP5) on cell proliferation and migration in human corneal endothelial (HCEC) and human corneal epithelial (CEPI17) cell lines, respectively.
  
  
  METHODS
  AQP1 and AQP5 were down regulated using siRNA following lipofectamine-mediated transfection in corneal endothelial and epithelial cells, respectively. Down-regulation was confirmed using RT-PCR, indirect immunofluorescence, and immunoblot analysis. Total internal reflection fluorescence (TIRF) microscopy was used to detect cell surface aquaporin expression. Cell proliferation was determined by SRB (sulfrodamine B) assay. Cell migration was determined by in vitro wound healing and migration assay.
  
  
  RESULTS
  In HCEC cells, AQP1 was localized to the cytosol as well as cell membrane and its down-regulation resulted in decreased cell proliferation and migration with a significant decrease in phosphorylated ERK (pERK). In CEPI17 cells AQP5 protein expression was also localized to cytosol as well as cell membrane. AQP5 down-regulation resulted in an increase in proliferation and cell migration with no significant difference in pERK.
  
  
  CONCLUSIONS
  AQP1 plays a role in HCEC proliferation and migration via the ERK signaling pathway and therefore may have significant implications in corneal endothelial dysfunction whereas; AQP5 may play an indirect role in human corneal epithelial cell proliferation and migration.
authors.affiliation: Department of Biomedical Sciences, Graduate School of Biomedical Sciences, University of North Texas Health Science Center, Fort Worth, TX 76107, USA.
authors.forename: Jwalitha
authors.initials: J
authors.lastname: Shankardas
authors.forename: Rajkumar V
authors.initials: RV
authors.lastname: Patil
authors.forename: Jamboor K
authors.initials: JK
authors.lastname: Vishwanatha
mesh_terms: D051398:Aquaporin 1 / Q000235:genetics* / Q000378:metabolism; D051402:Aquaporin 5 / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation; D003315:Cornea / Q000166:cytology*; D015536:Down-Regulation / Q000235:genetics*; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D004847:Epithelial Cells / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D020868:Gene Silencing; D006801:Humans; D007150:Immunohistochemistry; D010766:Phosphorylation; D021381:Protein Transport; D034741:RNA, Small Interfering / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: C496020:AQP1 protein, human; C496036:AQP5 protein, human; D051402:Aquaporin 5; D034741:RNA, Small Interfering; D051398:Aquaporin 1; D048049:Extracellular Signal-Regulated MAP Kinases
references.citation: Exp Cell Res. 1994 Oct;214(2):621-33
references.pmid: 7523155
references.citation: Int Rev Cytol. 2002;215:395-431
references.pmid: 11952236
references.citation: Nature. 2005 Apr 7;434(7034):786-92
references.pmid: 15815633
references.citation: Mol Vis. 2008 Apr 25;14:756-61
references.pmid: 18449356
references.citation: Prog Retin Eye Res. 2008 Jul;27(4):420-33
references.pmid: 18501660
references.citation: J Mol Med (Berl). 2008 May;86(5):523-9
references.pmid: 18311471
references.citation: J Intern Med. 2007 Jan;261(1):53-64
references.pmid: 17222168
references.citation: FASEB J. 2003 Aug;17(11):1508-10
references.pmid: 12824287
references.citation: J Histochem Cytochem. 2004 Oct;52(10):1341-50
references.pmid: 15385580
references.citation: Am J Physiol. 1998 May;274(5):C1332-45
references.pmid: 9612221
references.citation: Lab Invest. 2001 Feb;81(2):143-8
references.pmid: 11232635
references.citation: Invest Ophthalmol Vis Sci. 2003 Nov;44(11):4837-44
references.pmid: 14578406
references.citation: Biochem Biophys Res Commun. 2004 Dec 10;325(2):595-9
references.pmid: 15530434
references.citation: Nat Rev Mol Cell Biol. 2004 Sep;5(9):687-98
references.pmid: 15340377
references.citation: Expert Rev Mol Med. 2008 May 16;10:e13
references.pmid: 18482462
references.citation: Prog Retin Eye Res. 2003 May;22(3):359-89
references.pmid: 12852491
references.citation: Mol Vis. 2008;14:2604-15
references.pmid: 19122826
references.citation: Biochem Biophys Res Commun. 2009 Aug 28;386(3):483-7
references.pmid: 19539607
references.citation: PLoS One. 2009 Jul 08;4(7):e6167
references.pmid: 19584911
references.citation: Biochem Biophys Res Commun. 2008 Mar 7;367(2):291-8
references.pmid: 18155156
references.citation: Gynecol Oncol. 2006 Feb;100(2):294-9
references.pmid: 16242760
references.citation: J Immunol Methods. 1997 Oct 27;208(2):151-8
references.pmid: 9433470
references.citation: J Am Soc Nephrol. 2006 Jan;17(1):39-45
references.pmid: 16319186
references.citation: Pathologe. 2008 Nov;29 Suppl 2:357-62
references.pmid: 18841368
references.citation: Am J Physiol. 1998 Oct;275(4):C1151-7
references.pmid: 9755069
references.citation: PLoS One. 2008 Jul 09;3(7):e2594
references.pmid: 18612408
references.citation: J Biol Chem. 2002 May 24;277(21):19139-44
references.pmid: 11891232
references.citation: Exp Eye Res. 2009 Aug;89(2):159-65
references.pmid: 19298815
references.citation: J Neurochem. 2007 Jan;100(1):87-104
references.pmid: 17064359
references.citation: Invest Ophthalmol Vis Sci. 1999 May;40(6):1091-101
references.pmid: 10235542
references.citation: Invest Ophthalmol Vis Sci. 1995 Jan;36(1):32-40
references.pmid: 7822156
references.citation: Lancet. 2002 May 18;359(9319):1777-8; author reply 1778
references.pmid: 12049892
references.citation: Biochemistry. 2008 May 20;47(20):5657-67
references.pmid: 18426224
references.citation: Cancer Lett. 2008 Jun 8;264(1):54-62
references.pmid: 18423983
references.citation: J Membr Biol. 2006 Mar;210(2):105-15
references.pmid: 16868675
references.citation: Am J Pathol. 2008 Aug;173(2):518-25
references.pmid: 18583321
references.citation: Arthritis Rheum. 2003 Apr;48(4):1167-8
references.pmid: 12687572
references.citation: J Physiol Sci. 2009 Mar;59(2):113-22
references.pmid: 19340551
references.citation: Lancet. 2001 Mar 3;357(9257):688-9
references.pmid: 11247557
references.citation: Neuroscience. 2009 Jul 7;161(3):764-72
references.pmid: 19345723