Mitomycin C-induced cell death in mouse lens epithelial cells

Hyun Kyung Park, Kwang Won Lee, Jun S. Choi, Choun K. Joo

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The mode of action of mitomycin C (MMC)-induced cell death in lens epithelia was investigated using cell culture system. The cytotoxicity of MMC on α-TN4 mouse lens epithelial cells (LECs) was measured by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT)-based assay. To determine the type of cell death induced by MMC, we stained LECs with acridine orange and ethidium bromide, and performed TUNEL assay and electron microscopic examination. The activation of MMC was also investigated with N-acetly-L-cystein (NAC), a free radical scavenger, and dicumarol, an inhibitor of DT-diaphorase, to analyze one- and two-electron reduction pathways involved in the activation of MMC. MTT assay showed that 400 μg/ml of MMC decreased the cell viability of the control cells to 45%, and the cytotoxicity of MMC on α-TN4 cells was increased in a dose-dependent manner. Based on the results obtained from acridine orange and ethidium bromide staining, the TUNEL assay, and transmission electron microscopic examination, we confirmed that MMC-induced cell death, at 400 μg/ml of MMC, occurs with necrosis as well as apoptotis. The treatment of cells with 2 mMNAC for 1 h or 20 μM dicumarol for 30 min, prior to 5 min of MMC (400 μg/ml) treatment, restored the growth suppression to 76 and 80% of the control level, respectively. In addition, the cotreatment of cells with NAC and dicumarol restored cell viability to 90% of the control level. The cellular death in MMC-treated LECs showed the involvement of oxidative stress in this apoptosis accounting for 22% of the observed cell death at 400 μg/ml of MMC.

Original languageEnglish
Pages (from-to)213-219
Number of pages7
JournalOphthalmic Research
Volume34
Issue number4
DOIs
Publication statusPublished - 2002 Oct 17

Fingerprint

Mitomycin
Lenses
Cell Death
Epithelial Cells
Dicumarol
Electrons
Acridine Orange
Ethidium
In Situ Nick-End Labeling
Cell Survival
NAD(P)H Dehydrogenase (Quinone)
Free Radical Scavengers
Oxidative Stress
Necrosis
Epithelium
Cell Culture Techniques
Apoptosis
Staining and Labeling

Keywords

  • Apoptosis
  • Lens epithelial cells
  • Mitomycin C
  • Oxidative stress
  • Posterior capsule opacification

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Mitomycin C-induced cell death in mouse lens epithelial cells. / Park, Hyun Kyung; Lee, Kwang Won; Choi, Jun S.; Joo, Choun K.

In: Ophthalmic Research, Vol. 34, No. 4, 17.10.2002, p. 213-219.

Research output: Contribution to journalArticle

Park, Hyun Kyung ; Lee, Kwang Won ; Choi, Jun S. ; Joo, Choun K. / Mitomycin C-induced cell death in mouse lens epithelial cells. In: Ophthalmic Research. 2002 ; Vol. 34, No. 4. pp. 213-219.
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