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Phytosphingosine induces apoptotic cell death via caspase 8 activation and Bax translocation in human cancer cells.
Clin Cancer Res. 2003 Feb; 9(2):878-85.CC

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-G(1) 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.

Authors+Show Affiliations

Laboratory of Radiation Effect, Korea Cancer Center Hospital, Seoul 139-706.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

12576463

Citation

Park, Moon-Taek, et al. "Phytosphingosine Induces Apoptotic Cell Death Via Caspase 8 Activation and Bax Translocation in Human Cancer Cells." Clinical Cancer Research : an Official Journal of the American Association for Cancer Research, vol. 9, no. 2, 2003, pp. 878-85.
Park MT, Kang JA, Choi JA, et al. Phytosphingosine induces apoptotic cell death via caspase 8 activation and Bax translocation in human cancer cells. Clin Cancer Res. 2003;9(2):878-85.
Park, M. T., Kang, J. A., Choi, J. A., Kang, C. M., Kim, T. H., Bae, S., Kang, S., Kim, S., Choi, W. I., Cho, C. K., Chung, H. Y., Lee, Y. S., & Lee, S. J. (2003). Phytosphingosine induces apoptotic cell death via caspase 8 activation and Bax translocation in human cancer cells. Clinical Cancer Research : an Official Journal of the American Association for Cancer Research, 9(2), 878-85.
Park MT, et al. Phytosphingosine Induces Apoptotic Cell Death Via Caspase 8 Activation and Bax Translocation in Human Cancer Cells. Clin Cancer Res. 2003;9(2):878-85. PubMed PMID: 12576463.
* Article titles in AMA citation format should be in sentence-case
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-Mo, AU - Kim,Tae-Hwan, AU - Bae,Sangwoo, AU - Kang,Seongman, AU - Kim,Sujong, AU - Choi,Weon-Ik, AU - Cho,Chul-Koo, AU - Chung,Hee-Yong, AU - Lee,Yun-Sil, AU - Lee,Su-Jae, PY - 2003/2/11/pubmed PY - 2003/8/2/medline PY - 2003/2/11/entrez SP - 878 EP - 85 JF - Clinical cancer research : an official journal of the American Association for Cancer Research JO - Clin Cancer Res VL - 9 IS - 2 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-G(1) 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. SN - 1078-0432 UR - https://www.unboundmedicine.com/medline/citation/12576463/Phytosphingosine_induces_apoptotic_cell_death_via_caspase_8_activation_and_Bax_translocation_in_human_cancer_cells_ L2 - http://clincancerres.aacrjournals.org/cgi/pmidlookup?view=long&pmid=12576463 DB - PRIME DP - Unbound Medicine ER -