Korean J Ophthalmol.  1999 Jun;13(1):7-15. 10.3341/kjo.1999.13.1.7.

Mitomycin C-induced apoptosis in cultured human Tenon's capsule fibroblasts

• Kim JW
• Kim SK
• Song IH
• Kim IT

• KMID: 1081830
• DOI: http://doi.org/10.3341/kjo.1999.13.1.7

Abstract

To investigate the mitomycin C-induced apoptotic cell death of fibroblasts, the primarily cultured human Tenon's capsule fibroblasts were exposed to a clinically used dosage of 0.4 mg/ml of mitomycin C for 5 minutes. TUNEL (TdT-mediated dUTP-biotin nick end labeling) assay and electron microscopic studies were performed to determine the extent of mitomycin C-induced apoptosis. A flow cytometric study was performed to quantify the apoptotic cell population over time. The TUNEL stains were positive and electron microscopy showed features of apoptotic cell death in some fibroblasts 3 and 5 days after treatment. Flow cytometric analysis using Annexin V-propidium iodide double staining detected apoptotic cells 3 days after treatment. These apoptotic cell populations increased at 4 days and were sustained for one week. This study revealed that the clinical effects of mitomycin C on fibroblasts may be mediated not only by antiproliferative but also apoptotic cell death to some degree. Therefore, the apoptotic cell death of fibroblasts induced by mitomycin C should be considered to properly understand the mechanism of wound healing after trabeculectomy with adjunctive mitomycin C.

Keyword

Apoptosis; Electron microscopy; Fibroblast; Flow cytometry; Mitomycin C; TUNEL

MeSH Terms

Antibiotics, Antineoplastic/pharmacology*
Apoptosis/drug effects*
Cells, Cultured
Comparative Study
Connective Tissue/pathology
Connective Tissue/drug effects*
Fibroblasts/ultrastructure
Fibroblasts/drug effects*
Flow Cytometry
Human
In Situ Nick-End Labeling
Male
Middle Age
Mitomycin/pharmacology*
Wound Healing/drug effects