@article {1364584,
	title = {Decline in DJ-1 and decreased nuclear translocation of Nrf2 in Fuchs endothelial corneal dystrophy},
	journal = {Invest Ophthalmol Vis Sci},
	volume = {53},
	number = {9},
	year = {2012},
	month = {2012 Aug 24},
	pages = {5806-13},
	abstract = {PURPOSE: This study sought to determine factors involved in nuclear factor erythroid 2-related factor 2 (Nrf2) regulation and their response to oxidative stress in Fuchs endothelial corneal dystrophy (FECD) and normal corneal endothelial cells (CECs). METHODS: FECD corneal buttons were obtained from transplantations and normal human corneas from tissue banks. Oxidative stress was induced by tert-butyl hydroperoxide (tBHP). Protein and mRNA levels of Nrf2, DJ-1, p53, and Kelch-like ECH-associated protein1 (Keap1) were investigated using Western blotting and real-time PCR. Immunoprecipitation was used to detect levels of oxidized DJ-1 protein and Cullin 3- (Cul3)-regulated degradation of DJ-1 in immortalized FECD (FECDi) and normal CEC (HCECi) cell lines. Nrf2 subcellular localization was assessed by immunocytochemistry. RESULTS: Nrf2 protein stabilizer, DJ-1, decreased significantly in FECD CECs compared with normal, whereas Nrf2 protein repressor, Keap1, was unchanged at baseline but increased under oxidative stress. Under oxidative stress, normal CECs upregulated DJ-1 protein synthesis, whereas FECD CECs did not. DJ-1 decline correlated with increased DJ-1 oxidative modification and carbonylation in FECDi as compared with HCECi. Increased labeling of immunoprecipitated DJ-1 protein with anti-Cul3 antibody indicated enhanced DJ-1 degradation in FECDi as compared with HCECi. Following tBHP treatment, Nrf2 translocated from cytoplasm to nuclei in normal CECs, whereas Nrf2 nuclear localization was not observed in FECD. CONCLUSIONS: Decreased levels of DJ-1 in FECD at baseline and under oxidative stress correlate with impaired Nrf2 nuclear translocation and heightened cell susceptibility to apoptosis. Targeting the DJ-1/Nrf2 axis could yield a mechanism to slow CEC degeneration in FECD.},
	keywords = {Aged, Blotting, Western, Cell Nucleus, Corneal Transplantation, Endothelium, Corneal, Female, Fuchs{\textquoteright} Endothelial Dystrophy, Humans, Immunohistochemistry, Intracellular Signaling Peptides and Proteins, Kelch-Like ECH-Associated Protein 1, Male, NF-E2-Related Factor 2, Oncogene Proteins, Oxidative Stress, Protein Deglycase DJ-1, Protein Transport, Real-Time Polymerase Chain Reaction, RNA, Messenger, tert-Butylhydroperoxide, Tissue Donors, Tumor Suppressor Protein p53, Up-Regulation},
	issn = {1552-5783},
	doi = {10.1167/iovs.12-10119},
	author = {Bitar, Maya S and Liu, Cailing and Ziaei, Alireza and Chen, Yuming and Schmedt, Thore and Jurkunas, Ula V}
}
