PMID- 22815636
OWN - NLM
STAT- MEDLINE
DCOM- 20121113
LR  - 20211021
IS  - 1090-0535 (Electronic)
IS  - 1090-0535 (Linking)
VI  - 18
DP  - 2012
TI  - Quiescent keratocytes fail to repair MMC induced DNA damage leading to the 
      long-term inhibition of myofibroblast differentiation and wound healing.
PG  - 1828-39
AB  - PURPOSE: The purpose of this study was to determine the acute and long-term 
      effects of mitomycin C (MMC) on quiescent rabbit corneal keratocytes regarding 
      cell proliferation, myofibroblast differentiation and DNA repair. METHODS: 
      Quiescent keratocytes cultured in serum-free media were exposed to various 
      concentrations of MMC and then treated with transforming growth factor-beta (TGFbeta). 
      DNA damage was evaluated in both cultured keratocytes and live rabbit eyes 
      following treatment with MMC. The long-term ability of quiescent keratocytes to 
      repair MMC induced damage in vivo was evaluated in rabbits treated with MMC 2 
      months before 100 mum deep lamellar keratectomy (LK) injury. RESULTS: MMC 
      significantly blocked TGFbeta-induced cell proliferation and myofibroblast 
      differentiation in cultured quiescent keratocytes and altered the transcriptional 
      regulation of macrophage chemotactic protein-1 (MCP-1) and alpha smooth muscle 
      actin (alphaSMA). MMC also induced phosphorylation of the nuclear histone marker of 
      DNA damage, gammaH2AX (a member of the H2A histone family), without induction of cell 
      cycle entry or immediate DNA repair measured by Comet assay. In live rabbits, 0.2 
      mg/ml MMC significantly induced gammaH2AX nuclear immunostaining (p<0.05) throughout 
      the cornea and corneas receiving 0.2 mg/ml MMC treatment 2 months before LK 
      injury showed complete absence of any corneal scarring. CONCLUSIONS: MMC induces 
      DNA damage to quiescent corneal keratocytes, which remains unrepaired, resulting 
      in abnormal cell replication and gene transcription that leads to long-term 
      effects on corneal repair. Overall these findings suggest that there may be 
      long-term and perhaps permanent consequences to the application of MMC as an 
      anti-fibrotic therapy.
FAU - Jester, James V
AU  - Jester JV
AD  - Gavin Herbert Eye Institute, University of California-Irvine, CA, USA. 
      jjester@uci.edu
FAU - Nien, Chyong Jy
AU  - Nien CJ
FAU - Vasiliou, Vasilis
AU  - Vasiliou V
FAU - Brown, Donald J
AU  - Brown DJ
LA  - eng
GR  - EY017963/EY/NEI NIH HHS/United States
GR  - R01 EY007348/EY/NEI NIH HHS/United States
GR  - R01 EY019719/EY/NEI NIH HHS/United States
GR  - R24 EY016663/EY/NEI NIH HHS/United States
GR  - EY016663/EY/NEI NIH HHS/United States
GR  - R01 EY017963/EY/NEI NIH HHS/United States
GR  - EY07348/EY/NEI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20120704
PL  - United States
TA  - Mol Vis
JT  - Molecular vision
JID - 9605351
RN  - 0 (Actins)
RN  - 0 (Chemokine CCL2)
RN  - 0 (Culture Media, Serum-Free)
RN  - 0 (Histones)
RN  - 0 (Transforming Growth Factor beta)
RN  - 50SG953SK6 (Mitomycin)
SB  - IM
MH  - Actins/genetics/metabolism
MH  - Animals
MH  - Cell Differentiation/drug effects
MH  - Cell Proliferation/drug effects
MH  - Cells, Cultured
MH  - Chemokine CCL2/genetics/metabolism
MH  - Cicatrix/prevention & control
MH  - Comet Assay
MH  - Cornea/*drug effects/metabolism
MH  - Corneal Injuries
MH  - Corneal Keratocytes/*drug effects/metabolism/pathology
MH  - Culture Media, Serum-Free
MH  - *DNA Damage
MH  - DNA Repair/drug effects
MH  - Histones/genetics/metabolism
MH  - Mitomycin/*pharmacology
MH  - Myofibroblasts/drug effects/metabolism/pathology
MH  - Rabbits
MH  - Transcription, Genetic/drug effects
MH  - Transforming Growth Factor beta/pharmacology
MH  - Wound Healing/*drug effects
PMC - PMC3398499
EDAT- 2012/07/21 06:00
MHDA- 2012/11/14 06:00
PMCR- 2012/01/01
CRDT- 2012/07/21 06:00
PHST- 2012/05/01 00:00 [received]
PHST- 2012/07/01 00:00 [accepted]
PHST- 2012/07/21 06:00 [entrez]
PHST- 2012/07/21 06:00 [pubmed]
PHST- 2012/11/14 06:00 [medline]
PHST- 2012/01/01 00:00 [pmc-release]
AID - 189 [pii]
AID - 2012MOLVIS0208 [pii]
PST - ppublish
SO  - Mol Vis. 2012;18:1828-39. Epub 2012 Jul 4.