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Ursolic acid differentially modulates apoptosis in skin melanoma and retinal pigment epithelial cells exposed to UV–VIS broadband radiation

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Abstract

The signaling pathways via mTOR (mammalian target of rapamycin) and AMPK (AMP-activated protein kinase) play key roles in transcription, translation and carcinogenesis, and may be activated by light exposure. These pathways can be modulated by naturally occurring compounds, such as the triterpenoid, ursolic acid (UA). Previously, the transcription factors p53 and NF-κB, which transactivate mitochondrial apoptosis-related genes, were shown to be differentially modulated by UA. UA-modulated apoptosis, following exposure to UV–VIS radiation (ultraviolet to visible light broadband radiation, hereafter abbreviated to UVR), is observed to correspond to differential levels of oxidative stress in retinal pigment epithelial (RPE) and skin melanoma (SM) cells. The cellular response to this phytochemical was characterized using western blot, flow cytometry, microscopy with reactive oxidative species probes MitoTracker and dihydroethidium, and membrane permeability assay. UA pretreatment potentiated cell cycle arrest and UVR-induced apoptosis selectively in SM cells while reducing photo-oxidative stress in the DNA of RPE cells presumably by antioxidant activity of UA. Mechanistically, the nuclear transportation of p65 and p53 was reduced by UA administration prior to UVR exposure while the levels of p65 and p53 nuclear transportation in SM cells were sustained at a substantially higher level. Finally, the mitochondrial functional assay showed that UVR induced the collapse of the mitochondrial membrane potential, and this effect was exacerbated by rapamycin or UA pretreatment in SM preferentially. These results were consistent with reduced proliferation observed in the clonogenic assay, indicating that UA treatment enhanced the phototoxicity of UVR, by modulating the activation of p53 and NF-κB and initiating a mitogenic response to optical radiation that triggered mitochondria-dependent apoptosis, particularly in skin melanoma cells. The study indicates that this compound has multiple actions with the potential for protecting normal cells while sensitizing skin melanoma cells to UV irradiation.

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Abbreviations

Ursolic acid:

UA

Retinal pigment epithelial:

RPE

Skin melanoma:

SM

Ultraviolet to visible light:

UV–VIS

Ultraviolet to visible light broadband radiation:

UVR

Dihydroethidium:

DHE

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Acknowledgments

This study was supported by the Julio C. Palmaz Pilot Research Grant, the National Science Foundation Partnerships for Research and Education in Materials (NSF-PREM), Grant No. DMR-0934218 in collaboration with Northwestern University MRSEC, and a CTRC Oppenheimer Multi-Investigator Research Grant. Data generated in the Flow Cytometry Shared Resource Facility was supported by UTHSCSA, NIH-NCI P30 CA54174 (CTRC at UTHSCSA) and UL1RR025767 (CTSA Grant). We thank the faculty members in the Departments of Ophthalmology and Radiology of the UT Health Science Center for their assistance and advice. Fluorescence images were generated in the Core Optical Imaging Facility which is supported by the UTHSCSA and NIH-NCI P30 CA54174 (CTRC at UTHSCSA).

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Authors and Affiliations

  1. Department of Radiology, University of Texas Health Science Center at San Antonio, San Antonio, TX, USA

    Yuan-Hao Lee

  2. Department of Cellular and Structural Biology, University of Texas Health Science Center at San Antonio, San Antonio, TX, USA

    Exing Wang

  3. Department of Ophthalmology, University of Texas Health Science Center at San Antonio, Mail Stop 6230, UT Health Science Center, 7703 Floyd Curl Drive, San Antonio, TX, 78229-3900, USA

    Neeru Kumar & Randolph D. Glickman

  4. Center for Biomedical Neuroscience, University of Texas Health Science Center at San Antonio, San Antonio, TX, USA

    Randolph D. Glickman

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  1. Yuan-Hao Lee
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  4. Randolph D. Glickman
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Correspondence to Randolph D. Glickman.

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Lee, YH., Wang, E., Kumar, N. et al. Ursolic acid differentially modulates apoptosis in skin melanoma and retinal pigment epithelial cells exposed to UV–VIS broadband radiation. Apoptosis 19, 816–828 (2014). https://doi.org/10.1007/s10495-013-0962-z

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