Protective role for bcl-2 in experimentally induced cell death of bovine corneal endothelial cells

Choun Ki Joo, Kyung Sun Cho, Hyun Eui Kim, Jun Sub Choi, Young Jun Oh

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

To characterize the pattern of cell death and to investigate the potential role of bcl-2 in a death paradigm of corneal endothelial cells, primary cultures of bovine corneal endothelial (BCEN) cells were first established and treated with 0.01-1 μM staurosporine, a nonspecific protein kinase inhibitor. The pattern of BCEN cell death induced by staurosporine was apoptotic in nature, characterized by shrinkage of the cytoplasmic membrane, nuclear condensation and DNA fragmentation. Cotreatment of BCEN cells with Z-VAD-fmk (a caspase inhibitor) but not cycloheximide (a protein synthesis inhibitor) prevented staurosporine-induced cell death. To investigate the potential role of bcl-2, BCEN cells were transferred with a eukaryotic expression vector containing anti-apoptotic bcl-2 cDNA and characterized by reverse transcription-polymerase chain reaction (RT-PCR; BCEN/bcl-2). As measured by the MTT reduction assay after treatment with staurosporine, the survival rate of BCEN/bcl-2 cells was 48.0 ± 4.8% compared to 7.4 ± 2.1% in control BCEN cells. As determined by light microscopy, apoptotic changes such as nuclear condensation and apoptotic bodies were largely attenuated in BCEN/bcl-2 cells after staurosporine treatment although arborization of processes and rounding up of the cell body were not affected by overexpression of bcl-2. These results suggest that staurosporine induces apoptosis in a cycloheximide-independent but caspase-dependent manner and bcl-2 acts as a negative regulator in staurosporine-induced apoptosis of BCEN cells.

Original languageEnglish
Pages (from-to)287-296
Number of pages10
JournalOphthalmic Research
Volume31
Issue number4
DOIs
Publication statusPublished - 1999 Jul 1

Fingerprint

Staurosporine
Cell Death
Endothelial Cells
Cycloheximide
Apoptosis
Polymerase Chain Reaction
Protein Synthesis Inhibitors
Caspase Inhibitors
DNA Fragmentation
Protein Kinase Inhibitors
Caspases
Reverse Transcription
Microscopy
Complementary DNA
Cell Culture Techniques
Cell Membrane
Light

All Science Journal Classification (ASJC) codes

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Joo, Choun Ki ; Cho, Kyung Sun ; Kim, Hyun Eui ; Choi, Jun Sub ; Oh, Young Jun. / Protective role for bcl-2 in experimentally induced cell death of bovine corneal endothelial cells. In: Ophthalmic Research. 1999 ; Vol. 31, No. 4. pp. 287-296.
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Protective role for bcl-2 in experimentally induced cell death of bovine corneal endothelial cells. / Joo, Choun Ki; Cho, Kyung Sun; Kim, Hyun Eui; Choi, Jun Sub; Oh, Young Jun.

In: Ophthalmic Research, Vol. 31, No. 4, 01.07.1999, p. 287-296.

Research output: Contribution to journalArticle

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