Irofulven (6-hydroxymethylacylfulvene, MGI 114) induces caspase 8 and 9-mediated apoptosis in human pancreatic adenocarcinoma cells.

Abstract

BACKGROUND:

Irofulven (MGI 114) is a novel, clinically active sesquiterpene whose mechanism of action is not fully understood. We sought to identify apoptotic effectors induced by this agent in human pancreatic cancer cells.

MATERIALS AND METHODS:

MTT assay was used to assess IC50-Apoptosis was quantitated by flow cytometry and DAPI staining. Caspase activation was identified by western blot analysis.

RESULTS:

Irofulven was cytotoxic against all pancreatic cancer cell lines tested (IC50 1-18 microM), and induced 10-fold (4%+/- 2, vs. 41% +/- 5) induction of apoptosis. Irofulven-treated cells also demonstrated PARP3 cleavage and DAPI staining. Apoptosis was reduced to baseline levels by Z-VAD-FMK, a broad-spectrum caspase inhibitor. Western blot analysis revealed that caspases-3, -7, -8, and -9 were activated by irofulven. Time course evaluation demonstrated that caspases-8 and -9 were the initial species activated.

CONCLUSION:

Our data demonstrate that the cytotoxicity of irofulven in human pancreatic carcinoma cell lines is mediated by caspase-induced apoptosis.

• Authors: W Wang, S J Waters, J R MacDonald, D D Von Hoff, W E Strodel, A R Miller
• Journal: Anticancer research (Anticancer Res) 2001 May-Jun Vol. 21 Issue 3B Pg. 1789-94 ISSN: 0250-7005 [Print] Greece
• PMID: 11497260 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Amino Acid Chloromethyl Ketones
• Antineoplastic Agents, Alkylating
• Coloring Agents
• Enzyme Inhibitors
• Indoles
• Sesquiterpenes
• Tetrazolium Salts
• Thiazoles
• benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone
• Propidium
• DAPI
• irofulven
• CASP8 protein, human
• CASP9 protein, human
• Caspase 8
• Caspase 9
• Caspases
• thiazolyl blue

Topics

• Amino Acid Chloromethyl Ketones (pharmacology)
• Antineoplastic Agents, Alkylating (pharmacology)
• Apoptosis
• Blotting, Western
• Caspase 8
• Caspase 9
• Caspases (metabolism)
• Coloring Agents (pharmacology)
• DNA Fragmentation
• Enzyme Inhibitors (pharmacology)
• Flow Cytometry
• Humans
• Immunoblotting
• In Situ Nick-End Labeling
• Indoles (pharmacology)
• Inhibitory Concentration 50
• Pancreatic Neoplasms (enzymology, pathology)
• Propidium (pharmacology)
• Sesquiterpenes (pharmacology)
• Tetrazolium Salts (pharmacology)
• Thiazoles (pharmacology)
• Tumor Cells, Cultured