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Apoptosis in keratocytes caused by mitomycin C
Tae-im Kim1, Hungwon Tchah, Seung-ah Lee
1Department of Ophthalmology, University of Ulsan, College of Medicine, Asan Medical Center, Seoul, Korea.
Investigative Ophthalmology & Visual Science
|April 26, 2003
Summary
Mitomycin C induces apoptosis in rabbit corneal keratocytes via the caspase pathway, specifically involving caspase-8 and caspase-9, which are linked to the mitochondrial pathway.
Area of Science:
- Ophthalmology
- Cell Biology
- Pharmacology
Background:
- Corneal stromal wound healing is a complex process involving keratocytes.
- Understanding the cellular mechanisms that modulate this healing is crucial for developing effective treatments.
- Mitomycin C is a drug with antiproliferative properties that has been used in ophthalmic procedures.
Purpose of the Study:
- To quantify the effect of mitomycin C on rabbit corneal keratocytes.
- To determine mitomycin C's potential in modulating corneal stromal wound healing.
- To investigate the specific cellular pathway through which mitomycin C exerts its effects.
Main Methods:
- Rabbit keratocytes were cultured and exposed to varying concentrations of mitomycin C.
- Apoptosis was assessed using Hoechst staining and flow cytometry with annexin V.
- Cell viability was measured using the lactate dehydrogenase (LDH) assay.
- Involvement of caspases and the mitochondrial pathway was investigated using specific inhibitors and cytochrome c immunostaining.
Main Results:
- Mitomycin C induced characteristic apoptotic changes in keratocytes, including nuclear fragmentation.
- LDH activity, an indicator of cell damage, increased significantly with mitomycin C exposure in a dose- and time-dependent manner.
- Inhibitors of caspase-8 and caspase-9 significantly reduced mitomycin C-induced LDH activity.
- Cytochrome c release into the cytoplasm was observed, indicating mitochondrial pathway activation.
Conclusions:
- Mitomycin C induces apoptosis, not necrosis, in cultured corneal keratocytes.
- The apoptotic process is mediated through the caspase pathway, involving caspase-8 and caspase-9.
- Mitochondrial pathways are implicated in mitomycin C-induced apoptosis of corneal keratocytes.
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