Irofulven induces apoptosis in breast cancer cells regardless of caspase-3 status

Maryanne C S Herzig, Huiyun Liang, Anne E. Johnson, Barbara Woynarowska, Jan M. Woynarowski

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Caspase-3 deficiency can limit the efficiency of pro-apoptotic anticancer treatments. Irofulven (hydroxymethylacylfulvene, HMAF, MGI 114, NSC 683863) is an antitumor drug, currently in a Phase III and multiple Phase II trials, which can differentiate between tumor and normal cells in apoptosis induction. This study investigated whether apoptosis induced by irofulven requires caspase-3. Irofulven action was compared in breast cancer cells differing in caspase-3 status: deficient MCF-7 cells and proficient MDA-MB-231 cells and in normal human mammary epithelial cells, HMEC. Irofulven induces significant, concentration and time-dependent apoptotic DNA fragmentation in breast cancer cell lines, regardless of caspase-3 status. After 12, 24 and 48 h incubation at 1 μM irofulven (∼3 × GI50), fragmented DNA comprised 3.7, 14.1 and 34.6% and 8.4, 12.6 and 20.3% of total DNA in MCF-7 and MDA-MB-231 cells, respectively. Cell viability (trypan blue exclusion) remained largely unaffected during the first 24 h but decreased markedly after 48 h, indicating secondary necrosis. Net losses in cell numbers were apparent at 48 h. Normal HMEC cells were refractory to 1 μM drug with only ∼3-9% fragmented DNA after 12-48 h, although apoptosis was observed at drug levels >3 μM. The broad-spectrum caspase inhibitor Z-VAD-fmk inhibited irofulven-induced apoptosis of all cell lines at 20 μM with nearly complete abrogation of apoptosis at 100 μM. Irofulven treatment resulted in marginal caspase-3 processing in MDA-MB-231 and HMEC cells. These results indicate that whereas the caspase cascade mediates irofulven- induced apoptosis, caspase-3 is dispensable (supported by NIH CA70091 and CA78706).

Original languageEnglish (US)
Pages (from-to)133-143
Number of pages11
JournalBreast Cancer Research and Treatment
Volume71
Issue number2
DOIs
StatePublished - 2002

Fingerprint

Caspase 3
Apoptosis
Breast Neoplasms
DNA
irofulven
Cell Line
Caspase Inhibitors
Trypan Blue
MCF-7 Cells
DNA Fragmentation
Caspases
Pharmaceutical Preparations
Antineoplastic Agents
Cell Survival
Breast
Necrosis
Cell Count
Epithelial Cells

Keywords

  • Apoptosis
  • Apoptosis resistance
  • Caspase-3
  • Irofulven
  • MCF-7 cells

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Herzig, M. C. S., Liang, H., Johnson, A. E., Woynarowska, B., & Woynarowski, J. M. (2002). Irofulven induces apoptosis in breast cancer cells regardless of caspase-3 status. Breast Cancer Research and Treatment, 71(2), 133-143. https://doi.org/10.1023/A:1013855615712

Irofulven induces apoptosis in breast cancer cells regardless of caspase-3 status. / Herzig, Maryanne C S; Liang, Huiyun; Johnson, Anne E.; Woynarowska, Barbara; Woynarowski, Jan M.

In: Breast Cancer Research and Treatment, Vol. 71, No. 2, 2002, p. 133-143.

Research output: Contribution to journalArticle

Herzig, MCS, Liang, H, Johnson, AE, Woynarowska, B & Woynarowski, JM 2002, 'Irofulven induces apoptosis in breast cancer cells regardless of caspase-3 status', Breast Cancer Research and Treatment, vol. 71, no. 2, pp. 133-143. https://doi.org/10.1023/A:1013855615712
Herzig, Maryanne C S ; Liang, Huiyun ; Johnson, Anne E. ; Woynarowska, Barbara ; Woynarowski, Jan M. / Irofulven induces apoptosis in breast cancer cells regardless of caspase-3 status. In: Breast Cancer Research and Treatment. 2002 ; Vol. 71, No. 2. pp. 133-143.
@article{33f77a03134843398fbfe0f87d52497f,
title = "Irofulven induces apoptosis in breast cancer cells regardless of caspase-3 status",
abstract = "Caspase-3 deficiency can limit the efficiency of pro-apoptotic anticancer treatments. Irofulven (hydroxymethylacylfulvene, HMAF, MGI 114, NSC 683863) is an antitumor drug, currently in a Phase III and multiple Phase II trials, which can differentiate between tumor and normal cells in apoptosis induction. This study investigated whether apoptosis induced by irofulven requires caspase-3. Irofulven action was compared in breast cancer cells differing in caspase-3 status: deficient MCF-7 cells and proficient MDA-MB-231 cells and in normal human mammary epithelial cells, HMEC. Irofulven induces significant, concentration and time-dependent apoptotic DNA fragmentation in breast cancer cell lines, regardless of caspase-3 status. After 12, 24 and 48 h incubation at 1 μM irofulven (∼3 × GI50), fragmented DNA comprised 3.7, 14.1 and 34.6{\%} and 8.4, 12.6 and 20.3{\%} of total DNA in MCF-7 and MDA-MB-231 cells, respectively. Cell viability (trypan blue exclusion) remained largely unaffected during the first 24 h but decreased markedly after 48 h, indicating secondary necrosis. Net losses in cell numbers were apparent at 48 h. Normal HMEC cells were refractory to 1 μM drug with only ∼3-9{\%} fragmented DNA after 12-48 h, although apoptosis was observed at drug levels >3 μM. The broad-spectrum caspase inhibitor Z-VAD-fmk inhibited irofulven-induced apoptosis of all cell lines at 20 μM with nearly complete abrogation of apoptosis at 100 μM. Irofulven treatment resulted in marginal caspase-3 processing in MDA-MB-231 and HMEC cells. These results indicate that whereas the caspase cascade mediates irofulven- induced apoptosis, caspase-3 is dispensable (supported by NIH CA70091 and CA78706).",
keywords = "Apoptosis, Apoptosis resistance, Caspase-3, Irofulven, MCF-7 cells",
author = "Herzig, {Maryanne C S} and Huiyun Liang and Johnson, {Anne E.} and Barbara Woynarowska and Woynarowski, {Jan M.}",
year = "2002",
doi = "10.1023/A:1013855615712",
language = "English (US)",
volume = "71",
pages = "133--143",
journal = "Breast Cancer Research and Treatment",
issn = "0167-6806",
publisher = "Springer New York",
number = "2",

}

TY - JOUR

T1 - Irofulven induces apoptosis in breast cancer cells regardless of caspase-3 status

AU - Herzig, Maryanne C S

AU - Liang, Huiyun

AU - Johnson, Anne E.

AU - Woynarowska, Barbara

AU - Woynarowski, Jan M.

PY - 2002

Y1 - 2002

N2 - Caspase-3 deficiency can limit the efficiency of pro-apoptotic anticancer treatments. Irofulven (hydroxymethylacylfulvene, HMAF, MGI 114, NSC 683863) is an antitumor drug, currently in a Phase III and multiple Phase II trials, which can differentiate between tumor and normal cells in apoptosis induction. This study investigated whether apoptosis induced by irofulven requires caspase-3. Irofulven action was compared in breast cancer cells differing in caspase-3 status: deficient MCF-7 cells and proficient MDA-MB-231 cells and in normal human mammary epithelial cells, HMEC. Irofulven induces significant, concentration and time-dependent apoptotic DNA fragmentation in breast cancer cell lines, regardless of caspase-3 status. After 12, 24 and 48 h incubation at 1 μM irofulven (∼3 × GI50), fragmented DNA comprised 3.7, 14.1 and 34.6% and 8.4, 12.6 and 20.3% of total DNA in MCF-7 and MDA-MB-231 cells, respectively. Cell viability (trypan blue exclusion) remained largely unaffected during the first 24 h but decreased markedly after 48 h, indicating secondary necrosis. Net losses in cell numbers were apparent at 48 h. Normal HMEC cells were refractory to 1 μM drug with only ∼3-9% fragmented DNA after 12-48 h, although apoptosis was observed at drug levels >3 μM. The broad-spectrum caspase inhibitor Z-VAD-fmk inhibited irofulven-induced apoptosis of all cell lines at 20 μM with nearly complete abrogation of apoptosis at 100 μM. Irofulven treatment resulted in marginal caspase-3 processing in MDA-MB-231 and HMEC cells. These results indicate that whereas the caspase cascade mediates irofulven- induced apoptosis, caspase-3 is dispensable (supported by NIH CA70091 and CA78706).

AB - Caspase-3 deficiency can limit the efficiency of pro-apoptotic anticancer treatments. Irofulven (hydroxymethylacylfulvene, HMAF, MGI 114, NSC 683863) is an antitumor drug, currently in a Phase III and multiple Phase II trials, which can differentiate between tumor and normal cells in apoptosis induction. This study investigated whether apoptosis induced by irofulven requires caspase-3. Irofulven action was compared in breast cancer cells differing in caspase-3 status: deficient MCF-7 cells and proficient MDA-MB-231 cells and in normal human mammary epithelial cells, HMEC. Irofulven induces significant, concentration and time-dependent apoptotic DNA fragmentation in breast cancer cell lines, regardless of caspase-3 status. After 12, 24 and 48 h incubation at 1 μM irofulven (∼3 × GI50), fragmented DNA comprised 3.7, 14.1 and 34.6% and 8.4, 12.6 and 20.3% of total DNA in MCF-7 and MDA-MB-231 cells, respectively. Cell viability (trypan blue exclusion) remained largely unaffected during the first 24 h but decreased markedly after 48 h, indicating secondary necrosis. Net losses in cell numbers were apparent at 48 h. Normal HMEC cells were refractory to 1 μM drug with only ∼3-9% fragmented DNA after 12-48 h, although apoptosis was observed at drug levels >3 μM. The broad-spectrum caspase inhibitor Z-VAD-fmk inhibited irofulven-induced apoptosis of all cell lines at 20 μM with nearly complete abrogation of apoptosis at 100 μM. Irofulven treatment resulted in marginal caspase-3 processing in MDA-MB-231 and HMEC cells. These results indicate that whereas the caspase cascade mediates irofulven- induced apoptosis, caspase-3 is dispensable (supported by NIH CA70091 and CA78706).

KW - Apoptosis

KW - Apoptosis resistance

KW - Caspase-3

KW - Irofulven

KW - MCF-7 cells

UR - http://www.scopus.com/inward/record.url?scp=0036169791&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036169791&partnerID=8YFLogxK

U2 - 10.1023/A:1013855615712

DO - 10.1023/A:1013855615712

M3 - Article

C2 - 11883439

AN - SCOPUS:0036169791

VL - 71

SP - 133

EP - 143

JO - Breast Cancer Research and Treatment

JF - Breast Cancer Research and Treatment

SN - 0167-6806

IS - 2

ER -