c-Myc down-regulation increases susceptibility to cisplatin through reactive oxygen species-mediated apoptosis in M14 human melanoma cells

Annamaria Biroccio, Barbara Benassi, Sarah Amodei, Chiara Gabellini, Donatella Del Bufalo, Gabriella Zupi

Research output: Contribution to journalArticle

66 Citations (Scopus)

Abstract

Our aim in this work was to define the role of c-Myc in the susceptibility to cisplatin [cis-diamminedichloroplatinum(II) (CDDP)] in human melanoma cells. Two M14 melanoma cell clones obtained by transfection and expressing six to ten times lower c-Myc protein levels than the parental cells and the control clone were employed. Analysis of survival curves demonstrates an increase in CDDP sensitivity in c-Myc low-expressing clones if compared with the control clone and the parental line. The enhanced sensitivity is unrelated to the impairment in enzymatic DNA repair activity. Cell cycle analysis demonstrates that although the control clone is able to completely recover from the CDDP-induced S-G2/M block, this arrest is prolonged in c-Myc low-expressing clones and a fraction of cells undergoes apoptosis. Although no changes in P53, Bax, Bcl-2, and Bcl-XL/S protein levels are observed, apoptosis is associated with the formation of reactive oxygen species (ROS), activation of caspase-1, caspase-3 and cleavage of the specific caspase substrate poly-ADP-ribose polymerase. The use of the antioxidant N-acetyl cysteine and caspase inhibitors prevents CDDP-induced apoptosis in c-Myc low-expressing clones, demonstrating that ROS, caspase-1, and caspase-3 are required for apoptotic cell death. Moreover, ROS generation depends on caspase-1-like activation because the AcYVAD-cho inhibitor abrogates CDDP-induced ROS in the c-Myc low-expressing clones.

Original languageEnglish
Pages (from-to)174-182
Number of pages9
JournalMolecular Pharmacology
Volume60
Issue number1
Publication statusPublished - 2001

Fingerprint

Cisplatin
Melanoma
Reactive Oxygen Species
Down-Regulation
Clone Cells
Apoptosis
Caspase 1
Caspase 3
bcl-X Protein
Proto-Oncogene Proteins c-myc
Caspase Inhibitors
Poly(ADP-ribose) Polymerases
Survival Analysis
Caspases
DNA Repair
Transfection
Cysteine
Cell Cycle
Cell Death
Antioxidants

ASJC Scopus subject areas

  • Pharmacology

Cite this

c-Myc down-regulation increases susceptibility to cisplatin through reactive oxygen species-mediated apoptosis in M14 human melanoma cells. / Biroccio, Annamaria; Benassi, Barbara; Amodei, Sarah; Gabellini, Chiara; Del Bufalo, Donatella; Zupi, Gabriella.

In: Molecular Pharmacology, Vol. 60, No. 1, 2001, p. 174-182.

Research output: Contribution to journalArticle

@article{d4e32e337e0a410897c32e8dcd6f8a70,
title = "c-Myc down-regulation increases susceptibility to cisplatin through reactive oxygen species-mediated apoptosis in M14 human melanoma cells",
abstract = "Our aim in this work was to define the role of c-Myc in the susceptibility to cisplatin [cis-diamminedichloroplatinum(II) (CDDP)] in human melanoma cells. Two M14 melanoma cell clones obtained by transfection and expressing six to ten times lower c-Myc protein levels than the parental cells and the control clone were employed. Analysis of survival curves demonstrates an increase in CDDP sensitivity in c-Myc low-expressing clones if compared with the control clone and the parental line. The enhanced sensitivity is unrelated to the impairment in enzymatic DNA repair activity. Cell cycle analysis demonstrates that although the control clone is able to completely recover from the CDDP-induced S-G2/M block, this arrest is prolonged in c-Myc low-expressing clones and a fraction of cells undergoes apoptosis. Although no changes in P53, Bax, Bcl-2, and Bcl-XL/S protein levels are observed, apoptosis is associated with the formation of reactive oxygen species (ROS), activation of caspase-1, caspase-3 and cleavage of the specific caspase substrate poly-ADP-ribose polymerase. The use of the antioxidant N-acetyl cysteine and caspase inhibitors prevents CDDP-induced apoptosis in c-Myc low-expressing clones, demonstrating that ROS, caspase-1, and caspase-3 are required for apoptotic cell death. Moreover, ROS generation depends on caspase-1-like activation because the AcYVAD-cho inhibitor abrogates CDDP-induced ROS in the c-Myc low-expressing clones.",
author = "Annamaria Biroccio and Barbara Benassi and Sarah Amodei and Chiara Gabellini and {Del Bufalo}, Donatella and Gabriella Zupi",
year = "2001",
language = "English",
volume = "60",
pages = "174--182",
journal = "Molecular Pharmacology",
issn = "0026-895X",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "1",

}

TY - JOUR

T1 - c-Myc down-regulation increases susceptibility to cisplatin through reactive oxygen species-mediated apoptosis in M14 human melanoma cells

AU - Biroccio, Annamaria

AU - Benassi, Barbara

AU - Amodei, Sarah

AU - Gabellini, Chiara

AU - Del Bufalo, Donatella

AU - Zupi, Gabriella

PY - 2001

Y1 - 2001

N2 - Our aim in this work was to define the role of c-Myc in the susceptibility to cisplatin [cis-diamminedichloroplatinum(II) (CDDP)] in human melanoma cells. Two M14 melanoma cell clones obtained by transfection and expressing six to ten times lower c-Myc protein levels than the parental cells and the control clone were employed. Analysis of survival curves demonstrates an increase in CDDP sensitivity in c-Myc low-expressing clones if compared with the control clone and the parental line. The enhanced sensitivity is unrelated to the impairment in enzymatic DNA repair activity. Cell cycle analysis demonstrates that although the control clone is able to completely recover from the CDDP-induced S-G2/M block, this arrest is prolonged in c-Myc low-expressing clones and a fraction of cells undergoes apoptosis. Although no changes in P53, Bax, Bcl-2, and Bcl-XL/S protein levels are observed, apoptosis is associated with the formation of reactive oxygen species (ROS), activation of caspase-1, caspase-3 and cleavage of the specific caspase substrate poly-ADP-ribose polymerase. The use of the antioxidant N-acetyl cysteine and caspase inhibitors prevents CDDP-induced apoptosis in c-Myc low-expressing clones, demonstrating that ROS, caspase-1, and caspase-3 are required for apoptotic cell death. Moreover, ROS generation depends on caspase-1-like activation because the AcYVAD-cho inhibitor abrogates CDDP-induced ROS in the c-Myc low-expressing clones.

AB - Our aim in this work was to define the role of c-Myc in the susceptibility to cisplatin [cis-diamminedichloroplatinum(II) (CDDP)] in human melanoma cells. Two M14 melanoma cell clones obtained by transfection and expressing six to ten times lower c-Myc protein levels than the parental cells and the control clone were employed. Analysis of survival curves demonstrates an increase in CDDP sensitivity in c-Myc low-expressing clones if compared with the control clone and the parental line. The enhanced sensitivity is unrelated to the impairment in enzymatic DNA repair activity. Cell cycle analysis demonstrates that although the control clone is able to completely recover from the CDDP-induced S-G2/M block, this arrest is prolonged in c-Myc low-expressing clones and a fraction of cells undergoes apoptosis. Although no changes in P53, Bax, Bcl-2, and Bcl-XL/S protein levels are observed, apoptosis is associated with the formation of reactive oxygen species (ROS), activation of caspase-1, caspase-3 and cleavage of the specific caspase substrate poly-ADP-ribose polymerase. The use of the antioxidant N-acetyl cysteine and caspase inhibitors prevents CDDP-induced apoptosis in c-Myc low-expressing clones, demonstrating that ROS, caspase-1, and caspase-3 are required for apoptotic cell death. Moreover, ROS generation depends on caspase-1-like activation because the AcYVAD-cho inhibitor abrogates CDDP-induced ROS in the c-Myc low-expressing clones.

UR - http://www.scopus.com/inward/record.url?scp=0034948497&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034948497&partnerID=8YFLogxK

M3 - Article

C2 - 11408612

AN - SCOPUS:0034948497

VL - 60

SP - 174

EP - 182

JO - Molecular Pharmacology

JF - Molecular Pharmacology

SN - 0026-895X

IS - 1

ER -