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

Kim,  J W; Kim,  S K; Song,  I H; Kim,  I T

doi:1999.13.1.7

Korean J Ophthalmol > Volume 13(1); 1999 > Article

Comparative Study

Korean Journal of Ophthalmology 1999;13(1):7-15.
DOI: https://doi.org/10.3341/kjo.1999.13.1.7 Published online June 30, 1999.

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

J W Kim, S K Kim, I H Song, I T Kim

Department of Ophthalmology, School of Medicine, Catholic University of Taegu-Hyosung, Korea.

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.

Key Words: Apoptosis;Electron microscopy;Fibroblast;Flow cytometry;Mitomycin C;TUNEL