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Comparison of the Transcriptional Profiles of Melanocytes from Dark and Light Skinned Individuals under Basal Conditions and Following Ultraviolet-B Irradiation.

López S, Smith-Zubiaga I, García de Galdeano A, Boyano MD, García O, Gardeazábal J, Martinez-Cadenas C, Izagirre N, de la Rúa C, Alonso S - PLoS ONE (2015)

Bottom Line: Our results suggest that an interaction between ribosomal proteins and the P53 signaling pathway may occur in response to UVB in both DM and LM.Furthermore, the culture with KCM+ compared with KCM- had a noticeable effect on LM.This effect includes the activation of various signaling pathways such as the mTOR pathway, involved in the regulation of cell metabolism, growth, proliferation and survival.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetics, Physical Anthropology and Animal Physiology. University of the Basque Country UPV/EHU, Leioa, Bizkaia, Spain.

ABSTRACT
We analysed the whole-genome transcriptional profile of 6 cell lines of dark melanocytes (DM) and 6 of light melanocytes (LM) at basal conditions and after ultraviolet-B (UVB) radiation at different time points to investigate the mechanisms by which melanocytes protect human skin from the damaging effects of UVB. Further, we assessed the effect of different keratinocyte-conditioned media (KCM+ and KCM-) on melanocytes. Our results suggest that an interaction between ribosomal proteins and the P53 signaling pathway may occur in response to UVB in both DM and LM. We also observed that DM and LM show differentially expressed genes after irradiation, in particular at the first 6h after UVB. These are mainly associated with inflammatory reactions, cell survival or melanoma. Furthermore, the culture with KCM+ compared with KCM- had a noticeable effect on LM. This effect includes the activation of various signaling pathways such as the mTOR pathway, involved in the regulation of cell metabolism, growth, proliferation and survival. Finally, the comparison of the transcriptional profiles between LM and DM under basal conditions, and the application of natural selection tests in human populations allowed us to support the significant evolutionary role of MIF and ATP6V0B in the pigmentary phenotype.

No MeSH data available.


Related in: MedlinePlus

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Mentions: UV irradiation was performed in an ICH2 photoreactor (LuzChem, Canada) at 37°C. Cultures were irradiated at 75 mJ/cm2 UVB, based on our previous work [20], as we observed that this dosage led to a notable physiological effect but did not affect cell viability in both keratinocytes and melanocytes. Keratinocyte supernatants were harvested from both non-irradiated (KCM-) and irradiated keratinocytes (24 hours after treatment) (KCM+) and kept frozen at -80°C until subsequent use. Subconfluent melanocyte cultures were cultivated in Medium 254 supplemented with HMGS and KCM+ or KCM- medium in a proportion 1:1. The following day they were irradiated with 75 mJ/cm2 of UVB, and harvested at 6, 12 and 24 hours post irradiation. We used non-irradiated control cultures that were covered by aluminium foil during irradiation (Fig 1).


Comparison of the Transcriptional Profiles of Melanocytes from Dark and Light Skinned Individuals under Basal Conditions and Following Ultraviolet-B Irradiation.

López S, Smith-Zubiaga I, García de Galdeano A, Boyano MD, García O, Gardeazábal J, Martinez-Cadenas C, Izagirre N, de la Rúa C, Alonso S - PLoS ONE (2015)

Graphical scheme of the experimental design.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4526690&req=5

pone.0134911.g001: Graphical scheme of the experimental design.
Mentions: UV irradiation was performed in an ICH2 photoreactor (LuzChem, Canada) at 37°C. Cultures were irradiated at 75 mJ/cm2 UVB, based on our previous work [20], as we observed that this dosage led to a notable physiological effect but did not affect cell viability in both keratinocytes and melanocytes. Keratinocyte supernatants were harvested from both non-irradiated (KCM-) and irradiated keratinocytes (24 hours after treatment) (KCM+) and kept frozen at -80°C until subsequent use. Subconfluent melanocyte cultures were cultivated in Medium 254 supplemented with HMGS and KCM+ or KCM- medium in a proportion 1:1. The following day they were irradiated with 75 mJ/cm2 of UVB, and harvested at 6, 12 and 24 hours post irradiation. We used non-irradiated control cultures that were covered by aluminium foil during irradiation (Fig 1).

Bottom Line: Our results suggest that an interaction between ribosomal proteins and the P53 signaling pathway may occur in response to UVB in both DM and LM.Furthermore, the culture with KCM+ compared with KCM- had a noticeable effect on LM.This effect includes the activation of various signaling pathways such as the mTOR pathway, involved in the regulation of cell metabolism, growth, proliferation and survival.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetics, Physical Anthropology and Animal Physiology. University of the Basque Country UPV/EHU, Leioa, Bizkaia, Spain.

ABSTRACT
We analysed the whole-genome transcriptional profile of 6 cell lines of dark melanocytes (DM) and 6 of light melanocytes (LM) at basal conditions and after ultraviolet-B (UVB) radiation at different time points to investigate the mechanisms by which melanocytes protect human skin from the damaging effects of UVB. Further, we assessed the effect of different keratinocyte-conditioned media (KCM+ and KCM-) on melanocytes. Our results suggest that an interaction between ribosomal proteins and the P53 signaling pathway may occur in response to UVB in both DM and LM. We also observed that DM and LM show differentially expressed genes after irradiation, in particular at the first 6h after UVB. These are mainly associated with inflammatory reactions, cell survival or melanoma. Furthermore, the culture with KCM+ compared with KCM- had a noticeable effect on LM. This effect includes the activation of various signaling pathways such as the mTOR pathway, involved in the regulation of cell metabolism, growth, proliferation and survival. Finally, the comparison of the transcriptional profiles between LM and DM under basal conditions, and the application of natural selection tests in human populations allowed us to support the significant evolutionary role of MIF and ATP6V0B in the pigmentary phenotype.

No MeSH data available.


Related in: MedlinePlus