Role of promyelocytic leukemia zinc finger protein in proliferation of cultured human corneal endothelial cells.
OBJECTIVE
To review the role of promyelocytic leukemia zinc finger (PLZF), a transcriptional repressor and negative regulator of cell cycling, in the proliferation of cultured human corneal endothelial cells (HCECs).


METHODS
The expression pattern of PLZF mRNA was determined by reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time quantitative PCR in HCECs and normal human corneal epithelia. The effect of cell-cell contact on expression of the PLZF gene was studied after incubation of the cultured HCECs in EDTA. The proliferation rate of cultured HCECs was assayed by a real-time electronic sensing (RT-CES) system, and DNA microarray analysis was performed to find the PLZF-regulating genes in cultured HCECs infected with LacZ- and PLZF-carrying adenoviruses (Ad-LacZ, Ad-PLZF).


RESULTS
PLZF mRNA was expressed in HCECs in vivo and in completely confluent HCECs but not in subconfluent HCECs in vitro. Real-time PCR showed that the expression of PLZF mRNA was decreased by approximately 20-fold when incubated with EDTA and returned to a normal level as the cell-cell contact reformed. Cell proliferation assay by the RT-CES system showed that infection of cultured HCECs with Ad-PLZF inhibited proliferation.


CONCLUSIONS
These findings suggest that PLZF plays an important role in the suppression of proliferation of HCECs.
Shiraishi
Atsushi
A
Department of Ophthalmology, Ehime University Graduate School of Medicine, Shitsukawa, Toon, Ehime 791-0295, Japan. shiraia@m.ehime-u.ac.jp
Joko
Takeshi
T
Higashiyama
Shigeki
S
Ohashi
Yuichi
Y
D049109:Cell Proliferation; D002478:Cells, Cultured; D004728:Endothelium, Corneal / Q000166:cytology* / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D051741:Kruppel-Like Transcription Factors / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D000075722:Promyelocytic Leukemia Zinc Finger Protein; D012333:RNA, Messenger / Q000235:genetics*; D016335:Zinc Fingers
D051741:Kruppel-Like Transcription Factors; D000075722:Promyelocytic Leukemia Zinc Finger Protein; D012333:RNA, Messenger; C092037:ZBTB16 protein, human