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Live-cell imaging Unveils stimulus-specific dynamics of Nrf2 activation in UV-exposed melanoma cells: Implications
Nilar Win Htut1, Tasanee Onkoksoong2, Malinee Saelim2
1Department of Pharmacology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, 10700, Thailand; University of Medicine 2, Yangon, Khaymar Thi Rd, Yangon, Myanmar.
Abstract:
The transcription factor Nuclear factor e2-related factor 2 (Nrf2) is pivotal in orchestrating cellular antioxidant defense mechanisms, particularly in skin cells exposed to ultraviolet (UV) radiation and electrophilic phytochemicals. To comprehensively investigate Nrf2's role in maintaining cellular redox equilibrium following UV-induced stress, we engineered a novel Nrf2 fusion-based reporter system for real-time, live-cell quantification of Nrf2 activity in human melanoma cells. Utilizing live quantitative imaging, we dissected the kinetic profiles of Nrf2 activation in response to an array of stimuli, including UVA and UVB radiation, as well as a broad spectrum of phytochemicals including ferulic acid, gallic acid, hispidulin, p-coumaric acid, quercetin, resveratrol, tannic acid, and vanillic acid as well as well-known Nrf2 inducers, tert-butylhydroquinone (tBHQ) and sulforaphane (SFN). Intriguingly, we observed distinct dynamical patterns of Nrf2 activity contingent on the specific stimuli applied. Sustained activation of Nrf2 was empirically correlated with the increased antioxidant response element (ARE) activity. Our findings demonstrate the nuanced impact of different phenolic compounds on Nrf2 activity and the utility of our Nrf2-CTΔ16-YFP reporter in characterizing the dynamics of Nrf2 translocation in response to diverse stimuli. In summary, our innovative reporter system not only revealed compounds capable of modulating UVA-induced Nrf2 activity but also showcased its utility as a robust tool for future antioxidant compound screening efforts.
Insights
Nuclear factor e2-related factor 2 (Nrf2) plays a key role in skin cell defense against UV radiation. A new reporter system reveals distinct Nrf2 activation patterns by various compounds, aiding antioxidant research.
Area of Science:
- Cellular biology
- Molecular biology
- Dermatology
Background:
- Nuclear factor e2-related factor 2 (Nrf2) is crucial for cellular antioxidant defenses, especially in skin cells facing UV radiation.
- Understanding Nrf2's dynamic response to stressors is vital for maintaining redox balance.
Purpose of the Study:
- To develop and utilize a novel Nrf2 fusion-based reporter system for real-time, live-cell quantification of Nrf2 activity.
- To investigate the kinetic profiles of Nrf2 activation in human melanoma cells under various stimuli.
Main Methods:
- Engineered a novel Nrf2-CTΔ16-YFP fusion reporter system.
- Employed live quantitative imaging for real-time Nrf2 activity monitoring.
- Tested Nrf2 activation by UVA, UVB, phytochemicals, and known inducers like tBHQ and SFN.
Main Results:
- Observed distinct Nrf2 activity patterns depending on the specific stimulus.
- Demonstrated sustained Nrf2 activation correlated with increased antioxidant response element (ARE) activity.
- Identified specific phenolic compounds that modulate UVA-induced Nrf2 activity.
Conclusions:
- The Nrf2-CTΔ16-YFP reporter system effectively characterizes Nrf2 translocation dynamics.
- This system is a valuable tool for screening and identifying novel antioxidant compounds.
- Findings highlight the nuanced effects of phenolic compounds on Nrf2-mediated cellular responses.
