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The Effect of Lycopene Preexposure on UV-B-Irradiated Human Keratinocytes.

Ascenso A, Pedrosa T, Pinho S, Pinho F, de Oliveira JM, Cabral Marques H, Oliveira H, Simões S, Santos C - Oxid Med Cell Longev (2015)

Bottom Line: Lycopene did not significantly affect the profile of apoptotic, necrotic and viable cells in nonirradiated cells neither showed cytostatic effects.In irradiated cells, lycopene preexposure resulted in overexpression of BAX gene compared to nonexposed irradiated cells.This was accompanied by a cell cycle delay at S-phase transition and consequent decrease of cells in G0/G1 phase.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Investigação do Medicamento (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Avenida Professor Gama Pinto, 1649-003 Lisboa, Portugal ; Departamento de Biologia, Laboratório de Biotecnologia e Citómica, CESAM, Universidade de Aveiro, Campus Universitário de Santiago, 3810-193 Aveiro, Portugal.

ABSTRACT

Unlabelled: Lycopene has been reported as the antioxidant most quickly depleted in skin upon UV irradiation, and thus it might play a protective role. Our goal was to investigate the effects of preexposure to lycopene on UV-B-irradiated skin cells. Cells were exposed for 24 h to 10 M lycopene, and subsequently irradiated and left to recover for another 24 h period. Thereafter, several parameters were analyzed by FCM and

Rt-pcr: genotoxicity/clastogenicity by assessing the cell cycle distribution; apoptosis by performing the Annexin-V assay and analyzing gene expression of apoptosis biomarkers; and oxidative stress by ROS quantification. Lycopene did not significantly affect the profile of apoptotic, necrotic and viable cells in nonirradiated cells neither showed cytostatic effects. However, irradiated cells previously treated with lycopene showed an increase in both dead and viable subpopulations compared to nonexposed irradiated cells. In irradiated cells, lycopene preexposure resulted in overexpression of BAX gene compared to nonexposed irradiated cells. This was accompanied by a cell cycle delay at S-phase transition and consequent decrease of cells in G0/G1 phase. Thus, lycopene seems to play a corrective role in irradiated cells depending on the level of photodamage. Thus, our findings may have implications for the management of skin cancer.

No MeSH data available.


Related in: MedlinePlus

(a) Representative examples of Annexin V-FITC Dot-Plots Gating (FL1 LOG versus FITC LOG) of nonirradiated (NI) and UV-B (225 mJ/cm2) irradiated (IR) HaCaT cells not previously exposed or exposed to 10 μM complexed lycopene (Lyc-CD); (b) results are expressed as percentage (mean ± SD, n = 3) of nonapoptotic and viable cells, Q8: Annexin-FTIC (−) and PI (−); early apoptotic cells, Q5: Annexin-FTIC (+) and PI (−); predominantly late apoptotic or dead cells, Q6: Annexin- FTIC (+) and PI (+); and predominantly necrotic and dead cells, Q7: Annexin-FTIC (−) and PI (+). Statistical analysis: One-Way ANOVA with All Pairwise Multiple Comparison Procedures (Holm-Sidak method): means with different letters are significantly different (P < 0.05).
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fig6: (a) Representative examples of Annexin V-FITC Dot-Plots Gating (FL1 LOG versus FITC LOG) of nonirradiated (NI) and UV-B (225 mJ/cm2) irradiated (IR) HaCaT cells not previously exposed or exposed to 10 μM complexed lycopene (Lyc-CD); (b) results are expressed as percentage (mean ± SD, n = 3) of nonapoptotic and viable cells, Q8: Annexin-FTIC (−) and PI (−); early apoptotic cells, Q5: Annexin-FTIC (+) and PI (−); predominantly late apoptotic or dead cells, Q6: Annexin- FTIC (+) and PI (+); and predominantly necrotic and dead cells, Q7: Annexin-FTIC (−) and PI (+). Statistical analysis: One-Way ANOVA with All Pairwise Multiple Comparison Procedures (Holm-Sidak method): means with different letters are significantly different (P < 0.05).

Mentions: Annexin V assay differentiates subpopulations that are necrotic, apoptotic, or viable. Comparative analysis of these subpopulations in irradiated (225 mJ/cm2) and nonirradiated HaCaT cells, treated with 10 μM complexed lycopene, was performed using the Annexin V assay (Figure 6).


The Effect of Lycopene Preexposure on UV-B-Irradiated Human Keratinocytes.

Ascenso A, Pedrosa T, Pinho S, Pinho F, de Oliveira JM, Cabral Marques H, Oliveira H, Simões S, Santos C - Oxid Med Cell Longev (2015)

(a) Representative examples of Annexin V-FITC Dot-Plots Gating (FL1 LOG versus FITC LOG) of nonirradiated (NI) and UV-B (225 mJ/cm2) irradiated (IR) HaCaT cells not previously exposed or exposed to 10 μM complexed lycopene (Lyc-CD); (b) results are expressed as percentage (mean ± SD, n = 3) of nonapoptotic and viable cells, Q8: Annexin-FTIC (−) and PI (−); early apoptotic cells, Q5: Annexin-FTIC (+) and PI (−); predominantly late apoptotic or dead cells, Q6: Annexin- FTIC (+) and PI (+); and predominantly necrotic and dead cells, Q7: Annexin-FTIC (−) and PI (+). Statistical analysis: One-Way ANOVA with All Pairwise Multiple Comparison Procedures (Holm-Sidak method): means with different letters are significantly different (P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4664803&req=5

fig6: (a) Representative examples of Annexin V-FITC Dot-Plots Gating (FL1 LOG versus FITC LOG) of nonirradiated (NI) and UV-B (225 mJ/cm2) irradiated (IR) HaCaT cells not previously exposed or exposed to 10 μM complexed lycopene (Lyc-CD); (b) results are expressed as percentage (mean ± SD, n = 3) of nonapoptotic and viable cells, Q8: Annexin-FTIC (−) and PI (−); early apoptotic cells, Q5: Annexin-FTIC (+) and PI (−); predominantly late apoptotic or dead cells, Q6: Annexin- FTIC (+) and PI (+); and predominantly necrotic and dead cells, Q7: Annexin-FTIC (−) and PI (+). Statistical analysis: One-Way ANOVA with All Pairwise Multiple Comparison Procedures (Holm-Sidak method): means with different letters are significantly different (P < 0.05).
Mentions: Annexin V assay differentiates subpopulations that are necrotic, apoptotic, or viable. Comparative analysis of these subpopulations in irradiated (225 mJ/cm2) and nonirradiated HaCaT cells, treated with 10 μM complexed lycopene, was performed using the Annexin V assay (Figure 6).

Bottom Line: Lycopene did not significantly affect the profile of apoptotic, necrotic and viable cells in nonirradiated cells neither showed cytostatic effects.In irradiated cells, lycopene preexposure resulted in overexpression of BAX gene compared to nonexposed irradiated cells.This was accompanied by a cell cycle delay at S-phase transition and consequent decrease of cells in G0/G1 phase.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Investigação do Medicamento (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Avenida Professor Gama Pinto, 1649-003 Lisboa, Portugal ; Departamento de Biologia, Laboratório de Biotecnologia e Citómica, CESAM, Universidade de Aveiro, Campus Universitário de Santiago, 3810-193 Aveiro, Portugal.

ABSTRACT

Unlabelled: Lycopene has been reported as the antioxidant most quickly depleted in skin upon UV irradiation, and thus it might play a protective role. Our goal was to investigate the effects of preexposure to lycopene on UV-B-irradiated skin cells. Cells were exposed for 24 h to 10 M lycopene, and subsequently irradiated and left to recover for another 24 h period. Thereafter, several parameters were analyzed by FCM and

Rt-pcr: genotoxicity/clastogenicity by assessing the cell cycle distribution; apoptosis by performing the Annexin-V assay and analyzing gene expression of apoptosis biomarkers; and oxidative stress by ROS quantification. Lycopene did not significantly affect the profile of apoptotic, necrotic and viable cells in nonirradiated cells neither showed cytostatic effects. However, irradiated cells previously treated with lycopene showed an increase in both dead and viable subpopulations compared to nonexposed irradiated cells. In irradiated cells, lycopene preexposure resulted in overexpression of BAX gene compared to nonexposed irradiated cells. This was accompanied by a cell cycle delay at S-phase transition and consequent decrease of cells in G0/G1 phase. Thus, lycopene seems to play a corrective role in irradiated cells depending on the level of photodamage. Thus, our findings may have implications for the management of skin cancer.

No MeSH data available.


Related in: MedlinePlus