Activation of the Fas pathway independently of Fas ligand during apoptosis induced by camptothecin in p53 mutant human colon carcinoma cells

Rong Guang Shao, Chun X. Cao, Wilberto Nieves-Neira, M. T. Dimanche-Boitrel, Eric Solary, Yves Pommier

Research output: Contribution to journalArticle

78 Citations (Scopus)

Abstract

The present study explored the role of the cell surface receptor Fas (CD95/APO-1) in apoptosis induced by camptothecin (CPT) in the HT29 colon carcinoma cell line. CPT-induced apoptosis was associated with high molecular weight DNA fragmentation as measured by filter elution. This fragmentation was inhibited by the caspase inhibitor, z-VAD-fmk and by cycloheximide, which also prevented proteolytic activation of caspase-3 and poly(ADP-ribose)polymerase cleavage. Under such conditions, Fas, Fas ligand, Bax, and p21 expression were increased and Fas recruited the FADD adaptor. Fas expression increase was blocked by cycloheximide but not by z-VAD-fmk, consistent with caspase activation downstream from Fas. Treatment of HT29 cells with FasL or with the CH-11 agonistic anti-Fas antibody potentiated the apoptotic response of cells treated with CPT. The anti-Fas blocking antibody ZB4 and the Fas-ligand inhibitor failed to protect HT29 cells from CPT-induced apoptosis. Such a protection was obtained by transient expression of constructs encoding a dominant-negative mutant of FADD, FADD in an antisense orientation and E8 or MC159 viral proteins that inhibit Fas-induced apoptosis at the level of FADD and procaspase-8, respectively. Together, these data show that topoisomerase I-mediated DNA damage-induced apoptosis involves activation of the Fas pathway without detectable Fas-ligand requirement in CPT-treated cells.

Original languageEnglish (US)
Pages (from-to)1852-1859
Number of pages8
JournalOncogene
Volume20
Issue number15
DOIs
StatePublished - Apr 5 2001
Externally publishedYes

Fingerprint

Camptothecin
Fas Ligand Protein
Colon
Apoptosis
Carcinoma
HT29 Cells
Cycloheximide
Type I DNA Topoisomerase
Blocking Antibodies
Caspase Inhibitors
Poly(ADP-ribose) Polymerases
Caspase 8
Cell Surface Receptors
Viral Proteins
DNA Fragmentation
Caspases
Caspase 3
DNA Damage
Anti-Idiotypic Antibodies
Molecular Weight

Keywords

  • Bax
  • Caspase
  • FADD
  • Nuclease
  • Programmed cell death
  • z-VAD

ASJC Scopus subject areas

  • Cancer Research
  • Genetics
  • Molecular Biology

Cite this

Shao, R. G., Cao, C. X., Nieves-Neira, W., Dimanche-Boitrel, M. T., Solary, E., & Pommier, Y. (2001). Activation of the Fas pathway independently of Fas ligand during apoptosis induced by camptothecin in p53 mutant human colon carcinoma cells. Oncogene, 20(15), 1852-1859. https://doi.org/10.1038/sj.onc.1204264

Activation of the Fas pathway independently of Fas ligand during apoptosis induced by camptothecin in p53 mutant human colon carcinoma cells. / Shao, Rong Guang; Cao, Chun X.; Nieves-Neira, Wilberto; Dimanche-Boitrel, M. T.; Solary, Eric; Pommier, Yves.

In: Oncogene, Vol. 20, No. 15, 05.04.2001, p. 1852-1859.

Research output: Contribution to journalArticle

Shao, RG, Cao, CX, Nieves-Neira, W, Dimanche-Boitrel, MT, Solary, E & Pommier, Y 2001, 'Activation of the Fas pathway independently of Fas ligand during apoptosis induced by camptothecin in p53 mutant human colon carcinoma cells', Oncogene, vol. 20, no. 15, pp. 1852-1859. https://doi.org/10.1038/sj.onc.1204264
Shao, Rong Guang ; Cao, Chun X. ; Nieves-Neira, Wilberto ; Dimanche-Boitrel, M. T. ; Solary, Eric ; Pommier, Yves. / Activation of the Fas pathway independently of Fas ligand during apoptosis induced by camptothecin in p53 mutant human colon carcinoma cells. In: Oncogene. 2001 ; Vol. 20, No. 15. pp. 1852-1859.
@article{0cdd051891e1464880d4d6b91e55ceeb,
title = "Activation of the Fas pathway independently of Fas ligand during apoptosis induced by camptothecin in p53 mutant human colon carcinoma cells",
abstract = "The present study explored the role of the cell surface receptor Fas (CD95/APO-1) in apoptosis induced by camptothecin (CPT) in the HT29 colon carcinoma cell line. CPT-induced apoptosis was associated with high molecular weight DNA fragmentation as measured by filter elution. This fragmentation was inhibited by the caspase inhibitor, z-VAD-fmk and by cycloheximide, which also prevented proteolytic activation of caspase-3 and poly(ADP-ribose)polymerase cleavage. Under such conditions, Fas, Fas ligand, Bax, and p21 expression were increased and Fas recruited the FADD adaptor. Fas expression increase was blocked by cycloheximide but not by z-VAD-fmk, consistent with caspase activation downstream from Fas. Treatment of HT29 cells with FasL or with the CH-11 agonistic anti-Fas antibody potentiated the apoptotic response of cells treated with CPT. The anti-Fas blocking antibody ZB4 and the Fas-ligand inhibitor failed to protect HT29 cells from CPT-induced apoptosis. Such a protection was obtained by transient expression of constructs encoding a dominant-negative mutant of FADD, FADD in an antisense orientation and E8 or MC159 viral proteins that inhibit Fas-induced apoptosis at the level of FADD and procaspase-8, respectively. Together, these data show that topoisomerase I-mediated DNA damage-induced apoptosis involves activation of the Fas pathway without detectable Fas-ligand requirement in CPT-treated cells.",
keywords = "Bax, Caspase, FADD, Nuclease, Programmed cell death, z-VAD",
author = "Shao, {Rong Guang} and Cao, {Chun X.} and Wilberto Nieves-Neira and Dimanche-Boitrel, {M. T.} and Eric Solary and Yves Pommier",
year = "2001",
month = "4",
day = "5",
doi = "10.1038/sj.onc.1204264",
language = "English (US)",
volume = "20",
pages = "1852--1859",
journal = "Oncogene",
issn = "0950-9232",
publisher = "Nature Publishing Group",
number = "15",

}

TY - JOUR

T1 - Activation of the Fas pathway independently of Fas ligand during apoptosis induced by camptothecin in p53 mutant human colon carcinoma cells

AU - Shao, Rong Guang

AU - Cao, Chun X.

AU - Nieves-Neira, Wilberto

AU - Dimanche-Boitrel, M. T.

AU - Solary, Eric

AU - Pommier, Yves

PY - 2001/4/5

Y1 - 2001/4/5

N2 - The present study explored the role of the cell surface receptor Fas (CD95/APO-1) in apoptosis induced by camptothecin (CPT) in the HT29 colon carcinoma cell line. CPT-induced apoptosis was associated with high molecular weight DNA fragmentation as measured by filter elution. This fragmentation was inhibited by the caspase inhibitor, z-VAD-fmk and by cycloheximide, which also prevented proteolytic activation of caspase-3 and poly(ADP-ribose)polymerase cleavage. Under such conditions, Fas, Fas ligand, Bax, and p21 expression were increased and Fas recruited the FADD adaptor. Fas expression increase was blocked by cycloheximide but not by z-VAD-fmk, consistent with caspase activation downstream from Fas. Treatment of HT29 cells with FasL or with the CH-11 agonistic anti-Fas antibody potentiated the apoptotic response of cells treated with CPT. The anti-Fas blocking antibody ZB4 and the Fas-ligand inhibitor failed to protect HT29 cells from CPT-induced apoptosis. Such a protection was obtained by transient expression of constructs encoding a dominant-negative mutant of FADD, FADD in an antisense orientation and E8 or MC159 viral proteins that inhibit Fas-induced apoptosis at the level of FADD and procaspase-8, respectively. Together, these data show that topoisomerase I-mediated DNA damage-induced apoptosis involves activation of the Fas pathway without detectable Fas-ligand requirement in CPT-treated cells.

AB - The present study explored the role of the cell surface receptor Fas (CD95/APO-1) in apoptosis induced by camptothecin (CPT) in the HT29 colon carcinoma cell line. CPT-induced apoptosis was associated with high molecular weight DNA fragmentation as measured by filter elution. This fragmentation was inhibited by the caspase inhibitor, z-VAD-fmk and by cycloheximide, which also prevented proteolytic activation of caspase-3 and poly(ADP-ribose)polymerase cleavage. Under such conditions, Fas, Fas ligand, Bax, and p21 expression were increased and Fas recruited the FADD adaptor. Fas expression increase was blocked by cycloheximide but not by z-VAD-fmk, consistent with caspase activation downstream from Fas. Treatment of HT29 cells with FasL or with the CH-11 agonistic anti-Fas antibody potentiated the apoptotic response of cells treated with CPT. The anti-Fas blocking antibody ZB4 and the Fas-ligand inhibitor failed to protect HT29 cells from CPT-induced apoptosis. Such a protection was obtained by transient expression of constructs encoding a dominant-negative mutant of FADD, FADD in an antisense orientation and E8 or MC159 viral proteins that inhibit Fas-induced apoptosis at the level of FADD and procaspase-8, respectively. Together, these data show that topoisomerase I-mediated DNA damage-induced apoptosis involves activation of the Fas pathway without detectable Fas-ligand requirement in CPT-treated cells.

KW - Bax

KW - Caspase

KW - FADD

KW - Nuclease

KW - Programmed cell death

KW - z-VAD

UR - http://www.scopus.com/inward/record.url?scp=0035810046&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035810046&partnerID=8YFLogxK

U2 - 10.1038/sj.onc.1204264

DO - 10.1038/sj.onc.1204264

M3 - Article

C2 - 11313933

AN - SCOPUS:0035810046

VL - 20

SP - 1852

EP - 1859

JO - Oncogene

JF - Oncogene

SN - 0950-9232

IS - 15

ER -