Phytosphingosine induces apoptotic cell death via caspase 8 activation and Bax translocation in human cancer cells

Moon Taek Park, Jung A. Kang, Jung A. Choi, Chang M. Kang, Tae Hwan Kim, Sangwoo Bae, Seong Man Kang, Sujong Kim, Weon I. Choi, Chul K. Cho, Hee Yong Chung, Yun S. Lee, Su J. Lee

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

Purpose: Sphingolipid metabolites, such as sphingosine and ceramide, are highly bioactive compounds and are involved in diverse cell processes, including cell-cell interaction, cell proliferation, differentiation, and apoptosis. However, the physiological roles of phytosphingosine are poorly understood. In this study, we report that phytosphingosine can potently induce apoptotic cell death in human cancer cells via caspase activation and caspase-independent cytochrome c release. Experimental Design: Phytosphingosine-induced apoptosis was determined by Hoechst 33258 staining, flow cytometric analysis, and DNA fragmentation assay. Involvement of caspases was determined by immunoblot analysis and cell death detection assays after treatment with synthetic inhibitor z-Val-Ala-Asp-fluoromethyl ketone, z-DEVD-fmk, or z-IETD-fmk. Death receptor (DR) dependency was analyzed by examining expression of DRs (Fas, DR4, DR5, TNFR1, and R2), and interaction of Fas-associated death domain and caspase 8. Involvement of the mitochondria pathway was examined by monitoring of the mitochondria membrane potential, cytochrome c release, and Bax translocation. Results: Phytosphingosine-treated cells displayed several features of apoptosis, including increase of sub-G1 population, DNA fragmentation, and poly(ADP-ribose) polymerase cleavage. We observed that phytosphingosine cause activation of caspase 8 in a DR-independent fashion. Phytosphingosine also induced activation of caspase 9 and 3, loss of mitochondrial membrane potential, and the cytochrome c release from mitochondria. However, we failed to detect Bid cleavage. Moreover, caspase 8 inhibitor z-IETD-fmk did not affect phytosphingosine-induced cytochrome c release and caspase 9 activation, suggesting that phytosphingosine-induced cytochrome c release is caused by caspase 8-independent manner. Phytosphingosine induced mitochondrial translocation of Bax from the cytosol without changes in the protein levels of Bcl-2, Bcl-xL, and Bax. In addition, Bcl-2/Bax interaction was diminished after addition of phytosphingosine. Conclusion: These findings indicate that phytosphingosine induces apoptotic cell death in human cancer cells by direct activation of caspase 8, and by mitochondrial translocation of Bax and subsequent release of cytochrome c into cytoplasm, providing a potential mechanism for the anticancer activity of phytosphingosine.

Original languageEnglish
Pages (from-to)878-885
Number of pages8
JournalClinical Cancer Research
Volume9
Issue number2
Publication statusPublished - 2003 Feb 1

Fingerprint

phytosphingosine
Caspase 8
Cell Death
Cytochromes c
Neoplasms
Caspases
Death Domain Receptors
Mitochondria
Caspase 9
DNA Fragmentation
Apoptosis

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Park, M. T., Kang, J. A., Choi, J. A., Kang, C. M., Kim, T. H., Bae, S., ... Lee, S. J. (2003). Phytosphingosine induces apoptotic cell death via caspase 8 activation and Bax translocation in human cancer cells. Clinical Cancer Research, 9(2), 878-885.

Phytosphingosine induces apoptotic cell death via caspase 8 activation and Bax translocation in human cancer cells. / Park, Moon Taek; Kang, Jung A.; Choi, Jung A.; Kang, Chang M.; Kim, Tae Hwan; Bae, Sangwoo; Kang, Seong Man; Kim, Sujong; Choi, Weon I.; Cho, Chul K.; Chung, Hee Yong; Lee, Yun S.; Lee, Su J.

In: Clinical Cancer Research, Vol. 9, No. 2, 01.02.2003, p. 878-885.

Research output: Contribution to journalArticle

Park, MT, Kang, JA, Choi, JA, Kang, CM, Kim, TH, Bae, S, Kang, SM, Kim, S, Choi, WI, Cho, CK, Chung, HY, Lee, YS & Lee, SJ 2003, 'Phytosphingosine induces apoptotic cell death via caspase 8 activation and Bax translocation in human cancer cells', Clinical Cancer Research, vol. 9, no. 2, pp. 878-885.
Park, Moon Taek ; Kang, Jung A. ; Choi, Jung A. ; Kang, Chang M. ; Kim, Tae Hwan ; Bae, Sangwoo ; Kang, Seong Man ; Kim, Sujong ; Choi, Weon I. ; Cho, Chul K. ; Chung, Hee Yong ; Lee, Yun S. ; Lee, Su J. / Phytosphingosine induces apoptotic cell death via caspase 8 activation and Bax translocation in human cancer cells. In: Clinical Cancer Research. 2003 ; Vol. 9, No. 2. pp. 878-885.
@article{e4add5a4a2874302b5ff536133650a3f,
title = "Phytosphingosine induces apoptotic cell death via caspase 8 activation and Bax translocation in human cancer cells",
abstract = "Purpose: Sphingolipid metabolites, such as sphingosine and ceramide, are highly bioactive compounds and are involved in diverse cell processes, including cell-cell interaction, cell proliferation, differentiation, and apoptosis. However, the physiological roles of phytosphingosine are poorly understood. In this study, we report that phytosphingosine can potently induce apoptotic cell death in human cancer cells via caspase activation and caspase-independent cytochrome c release. Experimental Design: Phytosphingosine-induced apoptosis was determined by Hoechst 33258 staining, flow cytometric analysis, and DNA fragmentation assay. Involvement of caspases was determined by immunoblot analysis and cell death detection assays after treatment with synthetic inhibitor z-Val-Ala-Asp-fluoromethyl ketone, z-DEVD-fmk, or z-IETD-fmk. Death receptor (DR) dependency was analyzed by examining expression of DRs (Fas, DR4, DR5, TNFR1, and R2), and interaction of Fas-associated death domain and caspase 8. Involvement of the mitochondria pathway was examined by monitoring of the mitochondria membrane potential, cytochrome c release, and Bax translocation. Results: Phytosphingosine-treated cells displayed several features of apoptosis, including increase of sub-G1 population, DNA fragmentation, and poly(ADP-ribose) polymerase cleavage. We observed that phytosphingosine cause activation of caspase 8 in a DR-independent fashion. Phytosphingosine also induced activation of caspase 9 and 3, loss of mitochondrial membrane potential, and the cytochrome c release from mitochondria. However, we failed to detect Bid cleavage. Moreover, caspase 8 inhibitor z-IETD-fmk did not affect phytosphingosine-induced cytochrome c release and caspase 9 activation, suggesting that phytosphingosine-induced cytochrome c release is caused by caspase 8-independent manner. Phytosphingosine induced mitochondrial translocation of Bax from the cytosol without changes in the protein levels of Bcl-2, Bcl-xL, and Bax. In addition, Bcl-2/Bax interaction was diminished after addition of phytosphingosine. Conclusion: These findings indicate that phytosphingosine induces apoptotic cell death in human cancer cells by direct activation of caspase 8, and by mitochondrial translocation of Bax and subsequent release of cytochrome c into cytoplasm, providing a potential mechanism for the anticancer activity of phytosphingosine.",
author = "Park, {Moon Taek} and Kang, {Jung A.} and Choi, {Jung A.} and Kang, {Chang M.} and Kim, {Tae Hwan} and Sangwoo Bae and Kang, {Seong Man} and Sujong Kim and Choi, {Weon I.} and Cho, {Chul K.} and Chung, {Hee Yong} and Lee, {Yun S.} and Lee, {Su J.}",
year = "2003",
month = "2",
day = "1",
language = "English",
volume = "9",
pages = "878--885",
journal = "Clinical Cancer Research",
issn = "1078-0432",
publisher = "American Association for Cancer Research Inc.",
number = "2",

}

TY - JOUR

T1 - Phytosphingosine induces apoptotic cell death via caspase 8 activation and Bax translocation in human cancer cells

AU - Park, Moon Taek

AU - Kang, Jung A.

AU - Choi, Jung A.

AU - Kang, Chang M.

AU - Kim, Tae Hwan

AU - Bae, Sangwoo

AU - Kang, Seong Man

AU - Kim, Sujong

AU - Choi, Weon I.

AU - Cho, Chul K.

AU - Chung, Hee Yong

AU - Lee, Yun S.

AU - Lee, Su J.

PY - 2003/2/1

Y1 - 2003/2/1

N2 - Purpose: Sphingolipid metabolites, such as sphingosine and ceramide, are highly bioactive compounds and are involved in diverse cell processes, including cell-cell interaction, cell proliferation, differentiation, and apoptosis. However, the physiological roles of phytosphingosine are poorly understood. In this study, we report that phytosphingosine can potently induce apoptotic cell death in human cancer cells via caspase activation and caspase-independent cytochrome c release. Experimental Design: Phytosphingosine-induced apoptosis was determined by Hoechst 33258 staining, flow cytometric analysis, and DNA fragmentation assay. Involvement of caspases was determined by immunoblot analysis and cell death detection assays after treatment with synthetic inhibitor z-Val-Ala-Asp-fluoromethyl ketone, z-DEVD-fmk, or z-IETD-fmk. Death receptor (DR) dependency was analyzed by examining expression of DRs (Fas, DR4, DR5, TNFR1, and R2), and interaction of Fas-associated death domain and caspase 8. Involvement of the mitochondria pathway was examined by monitoring of the mitochondria membrane potential, cytochrome c release, and Bax translocation. Results: Phytosphingosine-treated cells displayed several features of apoptosis, including increase of sub-G1 population, DNA fragmentation, and poly(ADP-ribose) polymerase cleavage. We observed that phytosphingosine cause activation of caspase 8 in a DR-independent fashion. Phytosphingosine also induced activation of caspase 9 and 3, loss of mitochondrial membrane potential, and the cytochrome c release from mitochondria. However, we failed to detect Bid cleavage. Moreover, caspase 8 inhibitor z-IETD-fmk did not affect phytosphingosine-induced cytochrome c release and caspase 9 activation, suggesting that phytosphingosine-induced cytochrome c release is caused by caspase 8-independent manner. Phytosphingosine induced mitochondrial translocation of Bax from the cytosol without changes in the protein levels of Bcl-2, Bcl-xL, and Bax. In addition, Bcl-2/Bax interaction was diminished after addition of phytosphingosine. Conclusion: These findings indicate that phytosphingosine induces apoptotic cell death in human cancer cells by direct activation of caspase 8, and by mitochondrial translocation of Bax and subsequent release of cytochrome c into cytoplasm, providing a potential mechanism for the anticancer activity of phytosphingosine.

AB - Purpose: Sphingolipid metabolites, such as sphingosine and ceramide, are highly bioactive compounds and are involved in diverse cell processes, including cell-cell interaction, cell proliferation, differentiation, and apoptosis. However, the physiological roles of phytosphingosine are poorly understood. In this study, we report that phytosphingosine can potently induce apoptotic cell death in human cancer cells via caspase activation and caspase-independent cytochrome c release. Experimental Design: Phytosphingosine-induced apoptosis was determined by Hoechst 33258 staining, flow cytometric analysis, and DNA fragmentation assay. Involvement of caspases was determined by immunoblot analysis and cell death detection assays after treatment with synthetic inhibitor z-Val-Ala-Asp-fluoromethyl ketone, z-DEVD-fmk, or z-IETD-fmk. Death receptor (DR) dependency was analyzed by examining expression of DRs (Fas, DR4, DR5, TNFR1, and R2), and interaction of Fas-associated death domain and caspase 8. Involvement of the mitochondria pathway was examined by monitoring of the mitochondria membrane potential, cytochrome c release, and Bax translocation. Results: Phytosphingosine-treated cells displayed several features of apoptosis, including increase of sub-G1 population, DNA fragmentation, and poly(ADP-ribose) polymerase cleavage. We observed that phytosphingosine cause activation of caspase 8 in a DR-independent fashion. Phytosphingosine also induced activation of caspase 9 and 3, loss of mitochondrial membrane potential, and the cytochrome c release from mitochondria. However, we failed to detect Bid cleavage. Moreover, caspase 8 inhibitor z-IETD-fmk did not affect phytosphingosine-induced cytochrome c release and caspase 9 activation, suggesting that phytosphingosine-induced cytochrome c release is caused by caspase 8-independent manner. Phytosphingosine induced mitochondrial translocation of Bax from the cytosol without changes in the protein levels of Bcl-2, Bcl-xL, and Bax. In addition, Bcl-2/Bax interaction was diminished after addition of phytosphingosine. Conclusion: These findings indicate that phytosphingosine induces apoptotic cell death in human cancer cells by direct activation of caspase 8, and by mitochondrial translocation of Bax and subsequent release of cytochrome c into cytoplasm, providing a potential mechanism for the anticancer activity of phytosphingosine.

UR - http://www.scopus.com/inward/record.url?scp=0037314856&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037314856&partnerID=8YFLogxK

M3 - Article

C2 - 12576463

AN - SCOPUS:0037314856

VL - 9

SP - 878

EP - 885

JO - Clinical Cancer Research

JF - Clinical Cancer Research

SN - 1078-0432

IS - 2

ER -