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Signal transduction events elicited by cancer prevention compounds
1Department of Pharmaceutics and Pharmacodynamics, Center for Pharmaceutical Biotechnology, MC 870, College of Pharmacy, University of Illinois at Chicago, 900 S. Ashland Ave, Chicago, IL 60607, USA. kongt@cop.rutgers.edu
Abstract:
Many chemopreventive agents have been shown to modulate gene expression including induction of phase II detoxifying enzymes, such as glutathione S-transferases (GST) and quinone reductases (QR). Induction of phase II enzymes in general leads to protection of cells/tissues against exogenous and/or endogenous carcinogenic intermediates. The antioxidant or electrophile response element (ARE/EpRE) found at the 5'-flanking region of these phase II genes may play important role in mediating their induction by xenobiotics including chemopreventive agents. Members of the basic leucine zipper (bZIP) transcription factor, Nrf2 which heterodimerizes with Maf G/K, are found to bind to the ARE, and transcriptionally-activated ARE. Recently, we showed that the mitogen-activated protein kinases (MAPK) were activated by phase II gene inducers such as phenolic antioxidant butylated hydroxyanisol (BHA) and isothiocyanate sulforaphane (SUL), and involved in the transcription activation of ARE-mediated reporter gene. Transfection studies with wild-type and dominant negative mutants of Nrf2 and MAPK showed synergistic response during co-transfection as well as to phase II gene inducers. However, increasing the concentrations of these compounds such as BHA, the activities of cell death signaling molecules, caspases, were stimulated and resulted in apoptotic cell death. At these concentrations, BHA stimulated loss of mitochondrial membrane potential, cytochrome c release, and activation of caspase 3, 8 and 9 preceding apoptosis. Further increase in concentrations led to rapid cell necrosis. A model is proposed for BHA and SUL, in that at low concentrations, these potential chemopreventive agents may modulate MAPK pathway leading to transcription activation of Nrf2 and ARE with subsequent induction of cellular defensive enzymes including phase II detoxifying enzymes as well as other defensive genes, which may protect the cells against cellular injury, which is a homeostatic response. At higher concentrations, these agents may activate the caspase pathways, leading to apoptosis, a potential beneficial effect if occurs at preneoplastic/neoplastic tissues, but a potential cytotoxic response if occurs in normal tissues. On the other hand, some phenolic compounds such as resveratrol inhibits TPA- or UV-induced AP-1-mediated activity through the inhibition of c-Src non-receptor tyrosine kinase and MAPK pathways. It is possible that in proliferating or stimulated cells, these chemopreventive compounds may block proliferation by inhibiting these signaling kinases, whereas in non-proliferating or quiescent cells, some of these compounds may activate these signaling kinases leading to gene expression of cellular defensive enzymes such as phase II detoxifying enzymes. The studies of these and other signaling pathways may yield insights into the development of potential chemopreventive compounds.
Insights
Chemopreventive agents like BHA and SUL induce protective phase II detoxifying enzymes at low concentrations via the Nrf2-ARE pathway. However, higher concentrations trigger apoptosis and necrosis, suggesting a dose-dependent dual role in cellular defense and toxicity.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Chemopreventive agents often modulate gene expression, including the induction of phase II detoxifying enzymes like glutathione S-transferases (GST) and quinone reductases (QR).
- The antioxidant or electrophile response element (ARE/EpRE) is crucial for mediating the induction of these phase II genes by xenobiotics.
- The transcription factor Nrf2, in complex with Maf G/K, binds to the ARE and activates transcription.
Purpose of the Study:
- To investigate the role of mitogen-activated protein kinases (MAPK) in the transcriptional activation of ARE-mediated genes by chemopreventive agents.
- To elucidate the dose-dependent effects of chemopreventive agents on cellular signaling pathways, including cell death.
- To propose a model for the dual action of chemopreventive agents at different concentrations.
Main Methods:
- Transfection studies using wild-type and dominant-negative mutants of Nrf2 and MAPK.
- Assessment of caspase activity, mitochondrial membrane potential, and cytochrome c release.
- Dose-response studies with chemopreventive agents like butylated hydroxyanisole (BHA) and sulforaphane (SUL).
Main Results:
- MAPK activation by BHA and SUL is involved in the transcriptional activation of ARE-mediated reporter genes.
- Synergistic responses were observed between Nrf2 and MAPK during co-transfection and upon induction.
- At higher concentrations, BHA induced apoptosis via caspase activation and loss of mitochondrial membrane potential, followed by necrosis at even higher concentrations.
Conclusions:
- Low concentrations of BHA and SUL activate MAPK and Nrf2-ARE pathways, inducing cellular defense enzymes and protecting against injury.
- High concentrations of these agents can trigger apoptosis and necrosis, presenting a potential therapeutic window for chemoprevention but also a risk of cytotoxicity in normal tissues.
- Understanding these signaling pathways is key to developing effective chemopreventive compounds.