Elucidation of apoptosis induced by serum deprivation in cultured conjunctival epithelial cells

Akihiro Higuchi, Shigeto Shimmura, Tsutomu Takeuchi, M. Suematsu, Kazuo Tsubota

研究成果: Article

18 引用 (Scopus)

抄録

Background/aims: The conjunctival epithelial cell line, CCL20.2 (CCL), requires the presence of 10% fetal calf serum (FCS) in the medium to survive. To elucidate the molecular mechanism underlying such cell death, including the death signal for these cells, the activities of several caspases in the CCL were measured, and the effects of caspase inhibitors and serum components on cell death were examined. Methods: CCL was grown in Medium 199 containing 10% FCS, and the medium was changed to Medium 199 with or without 10% FCS, or medium without 10% FCS but containing caspase inhibitors or serum components. After 24 hours' incubation, the enzyme activities of caspases 1, 3, 8, and 9 in the culture supernatants were measured, and the effects of caspase inhibitors and serum components-for example, growth factors, lactoferrin, retinoic acid, were investigated. Results: DNA fragmentation was induced by serum deprivation, confirming that serum deprivation induces apoptosis in CCL. While the activities of caspases 3 and 8 were found to be increased, those of caspases 1 and 9 were not detected in the apoptotic cells. Z-VAD completely suppressed the caspase 3 activation, and specific inhibitors of caspases 1, 8, and 9 partially suppressed the activation. Serum deprivation induced a decrease in the cellular viability, which, however, partially recovered in the presence of caspase inhibitors, epidermal growth factor and retinoic acid. Conclusion: These results suggest that the apoptosis induced by serum deprivation involves caspases 1, 3, 8, and 9, and is suppressed by caspase inhibitors. EGF and retinoic acid have a key role in the maintenance of the ocular surface.

元の言語English
ページ(範囲)760-764
ページ数5
ジャーナルBritish Journal of Ophthalmology
90
発行部数6
DOI
出版物ステータスPublished - 2006 6

Fingerprint

Epithelial Cells
Apoptosis
Caspase Inhibitors
Serum
Caspase 1
Caspase 3
Tretinoin
Cell Death
Caspase 9
Caspase 8
Epidermal Growth Factor
Lactoferrin
DNA Fragmentation
Caspases
Intercellular Signaling Peptides and Proteins
Maintenance
Cell Line
Enzymes

ASJC Scopus subject areas

  • Ophthalmology

これを引用

@article{7a5ef7a711844393b8773e661cac5503,
title = "Elucidation of apoptosis induced by serum deprivation in cultured conjunctival epithelial cells",
abstract = "Background/aims: The conjunctival epithelial cell line, CCL20.2 (CCL), requires the presence of 10{\%} fetal calf serum (FCS) in the medium to survive. To elucidate the molecular mechanism underlying such cell death, including the death signal for these cells, the activities of several caspases in the CCL were measured, and the effects of caspase inhibitors and serum components on cell death were examined. Methods: CCL was grown in Medium 199 containing 10{\%} FCS, and the medium was changed to Medium 199 with or without 10{\%} FCS, or medium without 10{\%} FCS but containing caspase inhibitors or serum components. After 24 hours' incubation, the enzyme activities of caspases 1, 3, 8, and 9 in the culture supernatants were measured, and the effects of caspase inhibitors and serum components-for example, growth factors, lactoferrin, retinoic acid, were investigated. Results: DNA fragmentation was induced by serum deprivation, confirming that serum deprivation induces apoptosis in CCL. While the activities of caspases 3 and 8 were found to be increased, those of caspases 1 and 9 were not detected in the apoptotic cells. Z-VAD completely suppressed the caspase 3 activation, and specific inhibitors of caspases 1, 8, and 9 partially suppressed the activation. Serum deprivation induced a decrease in the cellular viability, which, however, partially recovered in the presence of caspase inhibitors, epidermal growth factor and retinoic acid. Conclusion: These results suggest that the apoptosis induced by serum deprivation involves caspases 1, 3, 8, and 9, and is suppressed by caspase inhibitors. EGF and retinoic acid have a key role in the maintenance of the ocular surface.",
author = "Akihiro Higuchi and Shigeto Shimmura and Tsutomu Takeuchi and M. Suematsu and Kazuo Tsubota",
year = "2006",
month = "6",
doi = "10.1136/bjo.2005.088203",
language = "English",
volume = "90",
pages = "760--764",
journal = "British Journal of Ophthalmology",
issn = "0007-1161",
publisher = "BMJ Publishing Group",
number = "6",

}

TY - JOUR

T1 - Elucidation of apoptosis induced by serum deprivation in cultured conjunctival epithelial cells

AU - Higuchi, Akihiro

AU - Shimmura, Shigeto

AU - Takeuchi, Tsutomu

AU - Suematsu, M.

AU - Tsubota, Kazuo

PY - 2006/6

Y1 - 2006/6

N2 - Background/aims: The conjunctival epithelial cell line, CCL20.2 (CCL), requires the presence of 10% fetal calf serum (FCS) in the medium to survive. To elucidate the molecular mechanism underlying such cell death, including the death signal for these cells, the activities of several caspases in the CCL were measured, and the effects of caspase inhibitors and serum components on cell death were examined. Methods: CCL was grown in Medium 199 containing 10% FCS, and the medium was changed to Medium 199 with or without 10% FCS, or medium without 10% FCS but containing caspase inhibitors or serum components. After 24 hours' incubation, the enzyme activities of caspases 1, 3, 8, and 9 in the culture supernatants were measured, and the effects of caspase inhibitors and serum components-for example, growth factors, lactoferrin, retinoic acid, were investigated. Results: DNA fragmentation was induced by serum deprivation, confirming that serum deprivation induces apoptosis in CCL. While the activities of caspases 3 and 8 were found to be increased, those of caspases 1 and 9 were not detected in the apoptotic cells. Z-VAD completely suppressed the caspase 3 activation, and specific inhibitors of caspases 1, 8, and 9 partially suppressed the activation. Serum deprivation induced a decrease in the cellular viability, which, however, partially recovered in the presence of caspase inhibitors, epidermal growth factor and retinoic acid. Conclusion: These results suggest that the apoptosis induced by serum deprivation involves caspases 1, 3, 8, and 9, and is suppressed by caspase inhibitors. EGF and retinoic acid have a key role in the maintenance of the ocular surface.

AB - Background/aims: The conjunctival epithelial cell line, CCL20.2 (CCL), requires the presence of 10% fetal calf serum (FCS) in the medium to survive. To elucidate the molecular mechanism underlying such cell death, including the death signal for these cells, the activities of several caspases in the CCL were measured, and the effects of caspase inhibitors and serum components on cell death were examined. Methods: CCL was grown in Medium 199 containing 10% FCS, and the medium was changed to Medium 199 with or without 10% FCS, or medium without 10% FCS but containing caspase inhibitors or serum components. After 24 hours' incubation, the enzyme activities of caspases 1, 3, 8, and 9 in the culture supernatants were measured, and the effects of caspase inhibitors and serum components-for example, growth factors, lactoferrin, retinoic acid, were investigated. Results: DNA fragmentation was induced by serum deprivation, confirming that serum deprivation induces apoptosis in CCL. While the activities of caspases 3 and 8 were found to be increased, those of caspases 1 and 9 were not detected in the apoptotic cells. Z-VAD completely suppressed the caspase 3 activation, and specific inhibitors of caspases 1, 8, and 9 partially suppressed the activation. Serum deprivation induced a decrease in the cellular viability, which, however, partially recovered in the presence of caspase inhibitors, epidermal growth factor and retinoic acid. Conclusion: These results suggest that the apoptosis induced by serum deprivation involves caspases 1, 3, 8, and 9, and is suppressed by caspase inhibitors. EGF and retinoic acid have a key role in the maintenance of the ocular surface.

UR - http://www.scopus.com/inward/record.url?scp=33744791137&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33744791137&partnerID=8YFLogxK

U2 - 10.1136/bjo.2005.088203

DO - 10.1136/bjo.2005.088203

M3 - Article

C2 - 16531423

AN - SCOPUS:33744791137

VL - 90

SP - 760

EP - 764

JO - British Journal of Ophthalmology

JF - British Journal of Ophthalmology

SN - 0007-1161

IS - 6

ER -