Abstract
Melanoma is one of the most chemoresistant cancers in patient care. The remission rate of current therapy remains low. DC-81, an antitumor antibiotic produced by Streptomyces species, belongs to pyrrolo[2,1-c][1,4]benzodiazepine (PBD), which is a potent inhibitor of nucleic acid synthesis. An enediyne contains either DNA intercalating groups or DNA minor groove binding functions and these are potent DNA-damaging agents due to their ability to generate benzenoid diradicals. We have previously reported an efficient synthesis and antitumor activity of a series of novel PBD hybrids linked with enediyne. The purpose of this study was to examine the mechanism of the antiproliferative effect of DC-81-enediyne agent on human melanoma A375 cells. DC-81-enediyne induced an increase in Ca2+ level and reactive oxygen species (ROS) generation as detected by flow cytometric assay. Western blot analysis showed that DC-81-enediyne induced the phosphorylation of p38 and activating transcription factor 2 (ATF-2). By using the luciferase reporter assay, activating protein-1 (AP-1) activity was further enhanced after A375 cells were treated with graded concentrations of DC-81-enediyne. DC-81-enediyne treatment-induced A375 cell apoptosis was significantly abrogated by the addition of Ca2+, ROS, and p38 inhibitors. Collectively, our studies indicate that DC-81-enediyne induces A375 cell apoptosis through an increased Ca2+ and ROS generation, which involves p38 phosphorylation and enhanced ATF-2/AP-1 expressions, leading to caspase-3 activity, poly(ADP-ribose)polymerase cleavage, M30 CytoDeath staining, and subsequent apoptotic cell death.
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Abbreviations
- AP-1:
-
Activating protein-1
- ATF-2:
-
Activating transcription factor 2
- DCFH-DA:
-
2′,7′-Dichlorofluorescein diacetate
- DMEM:
-
Dulbecco’s minimal essential medium
- ERK:
-
Extracellular signal-regulated kinase
- JNK:
-
c-Jun N-terminal kinase
- MAPK:
-
Mitogen-activated protein kinase
- PARP:
-
Poly(ADP-ribose)polymerase
- PBD:
-
Pyrrolo[2,1-c][1,4]benzodiazepine
- ROS:
-
Reactive oxygen species
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We thank the National Science Council of the Republic of China and the Kaohsiung Medical University Research Foundation for the financial support.
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Chen, CY., Chen, YK., Wang, JJ. et al. DC-81-enediyne induces apoptosis of human melanoma A375 cells: involvement of the ROS, p38 MAPK, and AP-1 signaling pathways. Cell Biol Toxicol 29, 85–99 (2013). https://doi.org/10.1007/s10565-012-9238-6
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DOI: https://doi.org/10.1007/s10565-012-9238-6