Age Associated Decrease of MT-1 Melatonin Receptor in Human Dermal Skin Fibroblasts Impairs Protection Against UV-Induced DNA Damage
Abstract
1. Introduction
2. Results
2.1. Melatonin Stimulates PER1 Clock Gene in Normal Human Dermal Fibroblast (NHDF) and in Normal Human Epidermal Keratinocytes (NHEK)
2.2. NHDF Express MT-1 Receptor and Its Level Is Decreased with Age
2.3. Characterization of MT-1 Receptor Knockdown Using siRNA Technology
2.4. Melatonin Receptor Knockdown Does not affect the Cell Viability of NHDF
2.5. MT-1 Receptor Knockdown Increases the Sensitivity of NHDF to UV Irradiation
2.6. Increased Oxidative Stress in MT-1 Receptor Knockdown NHDF
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. PER1 Gene Expression
4.3. MT-1 Melatonin Receptor Immunohistochemistry
4.4. Western Blotting
4.5. Electroporation of siRNA
4.6. Real-Time PCR
4.7. UV Exposure
4.8. Comet Assay (DNA Damage)
4.9. Cell Viability Measurement
4.10. H2O2 Measurement in UV-irradiated NHDF
4.11. Statistics
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
NHDF | Normal Human Dermal Fibroblasts |
NHEK | Normal Human Epidermal Keratinocytes |
ROS | Reactive Oxygen Species |
ANOVA | Analysis of Variance |
SEM | Standard Error of the Mean |
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Dong, K.; Goyarts, E.; Rella, A.; Pelle, E.; Wong, Y.H.; Pernodet, N. Age Associated Decrease of MT-1 Melatonin Receptor in Human Dermal Skin Fibroblasts Impairs Protection Against UV-Induced DNA Damage. Int. J. Mol. Sci. 2020, 21, 326. https://doi.org/10.3390/ijms21010326
Dong K, Goyarts E, Rella A, Pelle E, Wong YH, Pernodet N. Age Associated Decrease of MT-1 Melatonin Receptor in Human Dermal Skin Fibroblasts Impairs Protection Against UV-Induced DNA Damage. International Journal of Molecular Sciences. 2020; 21(1):326. https://doi.org/10.3390/ijms21010326
Chicago/Turabian StyleDong, Kelly, Earl Goyarts, Antonella Rella, Edward Pelle, Yung Hou Wong, and Nadine Pernodet. 2020. "Age Associated Decrease of MT-1 Melatonin Receptor in Human Dermal Skin Fibroblasts Impairs Protection Against UV-Induced DNA Damage" International Journal of Molecular Sciences 21, no. 1: 326. https://doi.org/10.3390/ijms21010326
APA StyleDong, K., Goyarts, E., Rella, A., Pelle, E., Wong, Y. H., & Pernodet, N. (2020). Age Associated Decrease of MT-1 Melatonin Receptor in Human Dermal Skin Fibroblasts Impairs Protection Against UV-Induced DNA Damage. International Journal of Molecular Sciences, 21(1), 326. https://doi.org/10.3390/ijms21010326