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M412458200v1
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Papers In Press, published online ahead of print January 19, 2005
J. Biol. Chem, 10.1074/jbc.M412458200
Submitted on November 3, 2004
Revised on January 14, 2005
Accepted on January 19, 2005

EGF-induced proliferation requires down-regulation of Pax6 in corneal epithelial cells

Tie Li and Luo Lu

Molecular Medicine, Harbor-UCLA Medical Center, Torrance, CA 90502

Corresponding Author: lluou{at}ucla.edu

Growth factors play important roles in regulating corneal epithelial cells proliferation/differentiation during wound healing. It is suggested that Pax6 involves corneal epithelium lineage-specific differentiation (1), however, the regulatory mechanism and function of Pax6 in growth factor-induced corneal epithelial responses is still unknown. In the present study, we found that the mitogenic effect of EGF in corneal epithelial cells required suppression of Pax6 activity through cellular mechanisms involving Erk signaling pathway-mediated increase in CTCF expression. EGF-induced CTCF activation subsequently inhibited pax6 expression by interacting with a CTCF-specific region upstream of Pax6 P0 promoter. Suppression of EGF-induced Erk activation by specific inhibitor or by the dominant expression of a silent Erk mutant effectively abolished the effects of EGF stimulation on regulations of CTCF and Pax6. Apparently, down-regulation of Pax6 expression induced by EGF is required for corneal epithelial proliferation because overexpression of Pax6 in these cells attenuated EGF-induced proliferation. In contrast, knockdown of mRNA expression with Pax6-specific or CTCF-specific siRNA in corneal epithelial cells significantly promoted or attenuated EGF-induced proliferation, respectively. Thus, our results revealed a new regulatory mechanism that involves cellular signaling events and Pax6 transcription regulation in growth factor-mediated proliferation. This suggests in corneal epithelial cells that inhibition of Pax6 expression is a prerequisite for EGF to elicit controls of cell growth and fate.


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