title: Microarray analysis of cell cycle gene expression in adult human corneal endothelial cells.
abstract: Corneal endothelial cells (ECs) form a monolayer that controls the hydration of the cornea and thus its transparency. Their almost nil proliferative status in humans is responsible, in several frequent diseases, for cell pool attrition that leads to irreversible corneal clouding. To screen for candidate genes involved in cell cycle arrest, we studied human ECs subjected to various environments thought to induce different proliferative profiles compared to ECs in vivo. Donor corneas (a few hours after death), organ-cultured (OC) corneas, in vitro confluent and non-confluent primary cultures, and an immortalized EC line were compared to healthy ECs retrieved in the first minutes of corneal grafts. Transcriptional profiles were compared using a cDNA array of 112 key genes of the cell cycle and analysed using Gene Ontology classification; cluster analysis and gene map presentation of the cell cycle regulation pathway were performed by GenMAPP. Results were validated using qRT-PCR on 11 selected genes. We found several transcripts of proteins implicated in cell cycle arrest and not previously reported in human ECs. Early G1-phase arrest effectors and multiple DNA damage-induced cell cycle arrest-associated transcripts were found in vivo and over-represented in OC and in vitro ECs. Though highly proliferative, immortalized ECs also exhibited overexpression of transcripts implicated in cell cycle arrest. These new effectors likely explain the stress-induced premature senescence that characterizes human adult ECs. They are potential targets for triggering and controlling EC proliferation with a view to increasing the cell pool of stored corneas or facilitating mass EC culture for bioengineered endothelial grafts.
authors.affiliation: Laboratory for corneal graft biology, engineering and imaging', EA2521, SFR143, Faculty of Medicine of the University Jean Monnet, Saint-Etienne, France.
authors.forename: Binh Minh
authors.initials: BM
authors.lastname: Ha Thi
authors.affiliation: Laboratory for corneal graft biology, engineering and imaging', EA2521, SFR143, Faculty of Medicine of the University Jean Monnet, Saint-Etienne, France; Department of Ophthalmology, University Hospital, Saint-Etienne, France.
authors.forename: Nelly
authors.initials: N
authors.lastname: Campolmi
authors.affiliation: Laboratory for corneal graft biology, engineering and imaging', EA2521, SFR143, Faculty of Medicine of the University Jean Monnet, Saint-Etienne, France.
authors.forename: Zhiguo
authors.initials: Z
authors.lastname: He
authors.affiliation: Laboratory for corneal graft biology, engineering and imaging', EA2521, SFR143, Faculty of Medicine of the University Jean Monnet, Saint-Etienne, France.
authors.forename: Aurélien
authors.initials: A
authors.lastname: Pipparelli
authors.affiliation: Laboratory for corneal graft biology, engineering and imaging', EA2521, SFR143, Faculty of Medicine of the University Jean Monnet, Saint-Etienne, France.
authors.forename: Chloé
authors.initials: C
authors.lastname: Manissolle
authors.affiliation: Service de Biologie Intégrative et de Génétique Moléculaire (SBIGeM), Institut de biologie et de technologies de Saclay (iBiTec-S), Commissariat à l'Energie Atomique, Gif-sur-Yvette, France.
authors.forename: Jean-Yves
authors.initials: JY
authors.lastname: Thuret
authors.affiliation: Laboratory for corneal graft biology, engineering and imaging', EA2521, SFR143, Faculty of Medicine of the University Jean Monnet, Saint-Etienne, France.
authors.forename: Simone
authors.initials: S
authors.lastname: Piselli
authors.affiliation: Laboratory for corneal graft biology, engineering and imaging', EA2521, SFR143, Faculty of Medicine of the University Jean Monnet, Saint-Etienne, France; Department of Pathology, University Hospital, Saint-Etienne, France.
authors.forename: Fabien
authors.initials: F
authors.lastname: Forest
authors.affiliation: Laboratory for corneal graft biology, engineering and imaging', EA2521, SFR143, Faculty of Medicine of the University Jean Monnet, Saint-Etienne, France; Department of Pathology, University Hospital, Saint-Etienne, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Peoc'h
authors.affiliation: Eye Bank, Etablissement Français du Sang Loire/Auvergne, Saint-Etienne, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Garraud
authors.affiliation: Laboratory for corneal graft biology, engineering and imaging', EA2521, SFR143, Faculty of Medicine of the University Jean Monnet, Saint-Etienne, France; Department of Ophthalmology, University Hospital, Saint-Etienne, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Gain
authors.affiliation: Laboratory for corneal graft biology, engineering and imaging', EA2521, SFR143, Faculty of Medicine of the University Jean Monnet, Saint-Etienne, France; Department of Ophthalmology, University Hospital, Saint-Etienne, France; Institut Universitaire de France, Paris, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Thuret
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D002453:Cell Cycle / Q000235:genetics*; D002455:Cell Division / Q000235:genetics; D002460:Cell Line; D016922:Cellular Senescence / Q000235:genetics; D018844:Cyclin-Dependent Kinases / Q000235:genetics; D016213:Cyclins / Q000235:genetics; D004728:Endothelium, Corneal / Q000166:cytology*; D020869:Gene Expression Profiling; D006801:Humans; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D009924:Organ Culture Techniques; D015398:Signal Transduction / Q000235:genetics
chemical_list: D016213:Cyclins; D018844:Cyclin-Dependent Kinases
references.citation: J Biol Chem. 2003 Mar 14;278(11):9754-60
references.pmid: 12519773
references.citation: Mol Cell. 2009 May 14;34(4):451-60
references.pmid: 19481525
references.citation: Int J Mol Med. 2006 Dec;18(6):1153-8
references.pmid: 17089020
references.citation: Nucleic Acids Res. 2012 Jan;40(Database issue):D109-14
references.pmid: 22080510
references.citation: Trans Am Ophthalmol Soc. 1953;51:669-786
references.pmid: 13216798
references.citation: Mol Cell Biochem. 2007 Dec;306(1-2):255-60
references.pmid: 17694278
references.citation: Oncol Rep. 2009 Sep;22(3):635-40
references.pmid: 19639215
references.citation: Invest Ophthalmol Vis Sci. 2006 Oct;47(10):4330-40
references.pmid: 17003423
references.citation: Invest Ophthalmol Vis Sci. 1996 Jul;37(8):1566-75
references.pmid: 8675399
references.citation: Mol Endocrinol. 2004 Dec;18(12):2895-907
references.pmid: 15358834
references.citation: DNA Repair (Amst). 2009 Aug 6;8(8):969-72
references.pmid: 19493704
references.citation: Mol Vis. 2007 Jul 23;13:1251-8
references.pmid: 17679950
references.citation: Invest Ophthalmol Vis Sci. 2009 May;50(5):2116-22
references.pmid: 19117931
references.citation: Invest Ophthalmol Vis Sci. 2004 Jun;45(6):1763-70
references.pmid: 15161838
references.citation: Mol Vis. 2011;17:3494-511
references.pmid: 22219645
references.citation: Mol Vis. 2005 May 06;11:328-34
references.pmid: 15889017
references.citation: Mol Carcinog. 2006 Mar;45(3):164-74
references.pmid: 16369997
references.citation: Nat Protoc. 2009;4(1):44-57
references.pmid: 19131956
references.citation: Exp Eye Res. 2004 Sep;79(3):367-76
references.pmid: 15336499
references.citation: Exp Eye Res. 2005 Dec;81(6):629-38
references.pmid: 16054624
references.citation: Acta Ophthalmol. 2010 Jun;88(4):413-9
references.pmid: 19604163
references.citation: Arch Ophthalmol. 2003 Feb;121(2):252-8
references.pmid: 12583793
references.citation: J Clin Invest. 1993 Apr;91(4):1713-20
references.pmid: 8386193
references.citation: Prog Retin Eye Res. 2003 May;22(3):359-89
references.pmid: 12852491
references.citation: PLoS One. 2011;6(12):e28310
references.pmid: 22194824
references.citation: Invest Ophthalmol Vis Sci. 2005 Oct;46(10):3597-603
references.pmid: 16186339
references.citation: PLoS One. 2012;7(1):e29677
references.pmid: 22276123
references.citation: Mol Cancer. 2004 Apr 26;3:12
references.pmid: 15109395
references.citation: Stem Cells. 2006 Apr;24(4):865-75
references.pmid: 16357341
references.citation: Invest Ophthalmol Vis Sci. 2006 Apr;47(4):1387-96
references.pmid: 16565372
references.citation: Cornea. 1991 Mar;10(2):136-48
references.pmid: 2019124
references.citation: Vision Res. 1970 Jan;10(1):107-8
references.pmid: 5435007
references.citation: Exp Eye Res. 2012 Feb;95(1):16-23
references.pmid: 21906590
references.citation: Exp Eye Res. 1984 Oct;39(4):497-504
references.pmid: 6499962
references.citation: Neuropathology. 2007 Aug;27(4):355-63
references.pmid: 17899689
references.citation: Oncogene. 1996 Sep 5;13(5):1103-9
references.pmid: 8806701
references.citation: Invest Ophthalmol Vis Sci. 2003 Apr;44(4):1521-8
references.pmid: 12657588
references.citation: Invest Ophthalmol Vis Sci. 2005 May;46(5):1626-31
references.pmid: 15851561
references.citation: Invest Ophthalmol Vis Sci. 2006 Nov;47(11):4803-9
references.pmid: 17065491
references.citation: Invest Ophthalmol Vis Sci. 1984 Mar;25(3):312-22
references.pmid: 6698749
references.citation: Br J Cancer. 2007 Jun 4;96(11):1723-8
references.pmid: 17486129
references.citation: Am J Med Genet A. 2007 Mar 1;143A(5):430-42
references.pmid: 17236194
references.citation: Curr Eye Res. 2012 Mar;37(3):179-86
references.pmid: 22335804
references.citation: Stem Cells Dev. 2013 Mar 1;22(5):828-39
references.pmid: 22974347
references.citation: Mol Vis. 2010 May 25;16:897-906
references.pmid: 20508865
references.citation: Tumour Biol. 2010 Jun;31(3):225-32
references.pmid: 20440596
references.citation: J Gen Virol. 2006 Mar;87(Pt 3):519-529
references.pmid: 16476973
references.citation: Stem Cells. 2012 Nov;30(11):2523-34
references.pmid: 22949402
references.citation: Invest Ophthalmol Vis Sci. 2000 Mar;41(3):660-7
references.pmid: 10711678
references.citation: Invest Ophthalmol Vis Sci. 2005 Oct;46(10):3604-15
references.pmid: 16186340
references.citation: Cells Tissues Organs. 2008;187(4):286-94
references.pmid: 18196893
references.citation: Plant Cell Rep. 2009 Apr;28(4):663-72
references.pmid: 19148648
references.citation: Genome Res. 2002 Feb;12(2):292-7
references.pmid: 11827948
references.citation: Biotechniques. 2005 Jul;39(1):75-85
references.pmid: 16060372
references.citation: Invest Ophthalmol Vis Sci. 1997 Mar;38(3):779-82
references.pmid: 9071233
references.citation: Stem Cells. 2010 Feb;28(2):308-17
references.pmid: 19967789
references.citation: Mol Vis. 2008 Jan 29;14:161-70
references.pmid: 18334933
references.citation: Exp Eye Res. 1988 Aug;47(2):205-15
references.pmid: 3409992
references.citation: Exp Eye Res. 2001 Apr;72(4):403-9
references.pmid: 11273668
references.citation: Exp Eye Res. 2012 Jul;100:40-51
references.pmid: 22575565
references.citation: Exp Eye Res. 2003 Oct;77(4):485-95
references.pmid: 12957147
references.citation: Cell. 2004 Sep 3;118(5):535-8
references.pmid: 15339658
references.citation: J Biomed Biotechnol. 2011;2011:412743
references.pmid: 22187525
references.citation: J Exp Clin Cancer Res. 2008 Sep 12;27:37
references.pmid: 18789152
references.citation: Mol Biol Cell. 2008 Nov;19(11):4968-79
references.pmid: 18784254
references.citation: Mol Vis. 2008 Aug 31;14:1589-99
references.pmid: 18769560
references.citation: Invest Ophthalmol Vis Sci. 1998 Dec;39(13):2572-83
references.pmid: 9856767
references.citation: Mol Vis. 2007 Oct 18;13:1984-2000
references.pmid: 17982423
references.citation: Rev Med Virol. 2002 Mar-Apr;12(2):81-92
references.pmid: 11921304
references.citation: Cornea. 2001 Oct;20(7):731-7
references.pmid: 11588426
references.citation: Acta Ophthalmol Scand. 2000 Apr;78(2):130-6
references.pmid: 10794243
references.citation: Invest Ophthalmol Vis Sci. 2004 Jun;45(6):1743-51
references.pmid: 15161835
references.citation: Invest Ophthalmol Vis Sci. 2003 Feb;44(2):594-9
references.pmid: 12556388
references.citation: Nucleic Acids Res. 2009 Jan;37(1):1-13
references.pmid: 19033363
references.citation: Oncogene. 1998 Dec 24;17(25):3247-59
references.pmid: 9916987
references.citation: Invest Ophthalmol Vis Sci. 1995 Jan;36(1):32-40
references.pmid: 7822156
references.citation: Invest Ophthalmol Vis Sci. 1993 May;34(6):2112-23
references.pmid: 8387979
references.citation: Invest Ophthalmol Vis Sci. 1996 Mar;37(4):645-55
references.pmid: 8595965
references.citation: J Surg Res. 2009 Dec;157(2):168-74
references.pmid: 19559447
references.citation: Exp Eye Res. 1986 Aug;43(2):251-8
references.pmid: 3758224
references.citation: Invest Ophthalmol Vis Sci. 2005 Nov;46(11):4086-91
references.pmid: 16249484
references.citation: Methods. 2001 Dec;25(4):402-8
references.pmid: 11846609
references.citation: Mol Cell Biol. 2003 Oct;23(19):6876-86
references.pmid: 12972606
references.citation: J Mol Med (Berl). 2006 Nov;84(11):901-10
references.pmid: 16972087
references.citation: Invest Ophthalmol Vis Sci. 2002 Jul;43(7):2152-9
references.pmid: 12091410
references.citation: Gene Ther. 2011 Aug;18(8):778-87
references.pmid: 21412281
references.citation: J Neurosci Methods. 2009 May 15;179(2):247-57
references.pmid: 19428534
references.citation: Invest Ophthalmol Vis Sci. 2009 Jun;50(6):2742-6
references.pmid: 19218607
references.citation: Aging Cell. 2012 Apr;11(2):234-40
references.pmid: 22128747
references.citation: Curr Eye Res. 2011 Dec;36(12):1116-22
references.pmid: 21999191
references.citation: Genes Cells. 2007 Nov;12(11):1289-300
references.pmid: 17986012
references.citation: Genes Dev. 1995 Mar 15;9(6):650-62
references.pmid: 7729684
references.citation: Nucleic Acids Res. 2000 Jan 1;28(1):27-30
references.pmid: 10592173
references.citation: BMC Bioinformatics. 2002 Jun 21;3:17
references.pmid: 12095422
references.citation: Cancer Res. 2002 Aug 1;62(15):4364-8
references.pmid: 12154041
references.citation: Tissue Eng Part A. 2011 Sep;17(17-18):2213-9
references.pmid: 21534849
references.citation: Invest Ophthalmol Vis Sci. 2000 Sep;41(10):2930-5
references.pmid: 10967047
references.citation: Arch Oral Biol. 2005 Aug;50(8):689-94
references.pmid: 15958200
references.citation: Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):2403-7
references.pmid: 7892279
references.citation: Am J Pathol. 2012 Jul;181(1):268-77
references.pmid: 22704232
references.citation: J Mol Endocrinol. 2002 Aug;29(1):23-39
references.pmid: 12200227
references.citation: Invest Ophthalmol Vis Sci. 2011 Mar 01;52(3):1641-9
references.pmid: 21087955
references.citation: Cornea. 2008 Feb;27(2):212-9
references.pmid: 18216579
references.citation: Cancer Res. 2002 Dec 15;62(24):7264-72
references.pmid: 12499268
references.citation: PLoS Biol. 2004 Feb;2(2):E7
references.pmid: 14737219
references.citation: J Cancer Res Clin Oncol. 2009 Jun;135(6):761-9
references.pmid: 19034516