5-Fluorouracil-Induced Apoptosis Changes in Cultured Corneal Epithelial Cells

Journal of Ocular Pharmacology and Therapeutics : the Official Journal of the Association for Ocular Pharmacology and Therapeutics
Kwou-Yeung WuShow-Jen Hong

Abstract

Repeated subconjunctival injections with 5-fluorouracil (5-FU) after trabeculectomy are used in glaucoma patients for the inhibition of overproliferation in wound site. Thus, a certain amount of the drug may penetrate into epithelial layer, where it causes toxicity to corneal epithelial cells. The aim of this study was to evaluate the toxic effects of 5-FU and mechanisms of drug-induced apoptosis in cultured corneal epithelial cells. Cellular damage and the caspase pathway were estimated with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The apoptotic characteristics were detected by flow cytometry, a TUNEL test, and western blotting in cultured corneal epithelial cells. The results indicated that 5-FU was toxic to corneal epithelial cells in a time- and dose-dependent manner. Pretreatment with a general caspase inhibitor (Z-VAD-FMK), a caspase-8 inhibitor (Z-IETD-FMK), and a caspase-9 inhibitor (Z-LEHD-FMK) reversed 5-FU-induced cellular damage. Following exposure to 5-FU, a flow cytometric assay with MitoLight dye demonstrated the significant loss of mitochondrial membrane potential. A positive TUNEL test revealed that cellular DNA apoptosis occurred following exposure to 0.5, 1, and 5 mg/mL 5-FU...Continue Reading

References

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Sep 27, 2001·Journal of Ocular Pharmacology and Therapeutics : the Official Journal of the Association for Ocular Pharmacology and Therapeutics·S J HongY H Lai
Oct 28, 2005·Current Eye Research·Kwou-Yeung WuShow-Jen Hong
Jan 18, 2006·Eye & Contact Lens·Danielle M RobertsonH Dwight Cavanagh
Oct 18, 2006·Graefe's Archive for Clinical and Experimental Ophthalmology = Albrecht Von Graefes Archiv Für Klinische Und Experimentelle Ophthalmologie·Eva Kristina ReinthalKarl Ulrich Bartz-Schmidt
Feb 13, 2008·Cell Biology and Toxicology·Jose A Aquino EsperanzaNora Cristina Brandan
Oct 7, 2008·Basic & Clinical Pharmacology & Toxicology·Su Jin KangHai Won Chung
Jun 15, 2011·Clinical & Experimental Ophthalmology·Justyna Jurkowska-DudzińskaTomasz Zarnowski
Jun 27, 2012·Journal of Ocular Pharmacology and Therapeutics : the Official Journal of the Association for Ocular Pharmacology and Therapeutics·Tulay SimsekUfuk Elgin

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Methods Mentioned

BETA
electrophoresis
flow cytometry
fluorescence microscopy
protein assay

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