Naringenin Protects HaCaT Human Keratinocytes Against UVB-induced Apoptosis and Enhances the Removal of Cyclobutane Pyrimidine Dimers from the Genome

Many naturally occurring agents are believed to protect against UV‐induced skin damage. In this study, we have investigated the effects of naringenin (NG), a naturally occurring citrus flavonone, on the removal of UVB‐induced cyclobutane pyrimidine dimers (CPD) from the genome and apoptosis in immor...

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Published in:Photochemistry and photobiology Vol. 84; no. 2; pp. 307 - 316
Main Authors: El-Mahdy, Mohamed A., Zhu, Qianzheng, Wang, Qi-En, Wani, Gulzar, Patnaik, Srinivas, Zhao, Qun, Arafa, El-Shaimaa, Barakat, Bassant, Mir, Safita N., Wani, Altaf A.
Format: Journal Article
Language:English
Published: Oxford, UK Blackwell Publishing Ltd 01.03.2008
Subjects:
Amino Acid Sequence
Apoptosis
Apoptosis - drug effects
Apoptosis - radiation effects
Biotechnology
Cell Line
Chemicals
Citrus
Deoxyribonucleic acid
DNA
DNA repair
Flavanones - pharmacology
Free radicals
Genetic disorders
Genome, Human
Human subjects
Humans
Keratinocytes - drug effects
Keratinocytes - radiation effects
Molecular Sequence Data
Monoclonal antibodies
Mortality
Pyrimidine Dimers - isolation & purification
Signal transduction
Ultraviolet Rays
ISSN:0031-8655, 1751-1097
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Abstract Many naturally occurring agents are believed to protect against UV‐induced skin damage. In this study, we have investigated the effects of naringenin (NG), a naturally occurring citrus flavonone, on the removal of UVB‐induced cyclobutane pyrimidine dimers (CPD) from the genome and apoptosis in immortalized p53‐mutant human keratinocyte HaCaT cells. The colony‐forming assay shows that treatment with NG significantly increases long‐term cell survival after UVB irradiation. NG treatment also protects the cells from UVB‐induced apoptosis, as indicated by the absence of the 180 base pair DNA ladders and the appearance of sub‐G1 peak using agarose gel electrophoresis and flow cytometric analysis, respectively. The UVB‐induced poly (ADP‐ribose) polymerase‐1 (PARP‐1) cleavage, caspase activation and Bax/Bcl2 ratio were modulated following NG treatment, indicating an antiapoptotic effect of NG in UVB‐damaged cells that occurs at least in part via caspase cascade pathway. Moreover, treatment of UVB‐irradiated HaCaT cells with NG enhances the removal of CPD from the genome, as observed by both direct quantitation of CPD in genomic DNA and immuno‐localization of the damage within the nuclei. The study provides a molecular basis for the action of NG as a promising natural flavonoid in preventing skin aging and carcinogenesis.
AbstractList Many naturally occurring agents are believed to protect against UV‐induced skin damage. In this study, we have investigated the effects of naringenin (NG), a naturally occurring citrus flavonone, on the removal of UVB‐induced cyclobutane pyrimidine dimers (CPD) from the genome and apoptosis in immortalized p53‐mutant human keratinocyte HaCaT cells. The colony‐forming assay shows that treatment with NG significantly increases long‐term cell survival after UVB irradiation. NG treatment also protects the cells from UVB‐induced apoptosis, as indicated by the absence of the 180 base pair DNA ladders and the appearance of sub‐G 1 peak using agarose gel electrophoresis and flow cytometric analysis, respectively. The UVB‐induced poly (ADP‐ribose) polymerase‐1 (PARP‐1) cleavage, caspase activation and Bax/Bcl2 ratio were modulated following NG treatment, indicating an antiapoptotic effect of NG in UVB‐damaged cells that occurs at least in part via caspase cascade pathway. Moreover, treatment of UVB‐irradiated HaCaT cells with NG enhances the removal of CPD from the genome, as observed by both direct quantitation of CPD in genomic DNA and immuno‐localization of the damage within the nuclei. The study provides a molecular basis for the action of NG as a promising natural flavonoid in preventing skin aging and carcinogenesis.
UV exposure also causes DNA damage including UV-induced CPD and 6-4PP and these adducts can be removed by nucleotide excision repair (NER). Monoclonal antibodies against actin and XPB were from Neomarkers (Fremont, CA) and Santa Cruz Biotechnology (Santa Cruz, CA), respectively.\n The fact that cyclin A-associated kinase activity is required for entry into S phase, completion of S phase and entry into M phase (61-63) suggests that cyclin/cdk complexes may have a role in DNA repair (64).
Many naturally occurring agents are believed to protect against UV-induced skin damage. In this study, we have investigated the effects of naringenin (NG), a naturally occurring citrus flavonone, on the removal of UVB-induced cyclobutane pyrimidine dimers (CPD) from the genome and apoptosis in immortalized p53-mutant human keratinocyte HaCaT cells. The colony-forming assay shows that treatment with NG significantly increases long-term cell survival after UVB irradiation. NG treatment also protects the cells from UVB-induced apoptosis, as indicated by the absence of the 180 base pair DNA ladders and the appearance of sub-G1 peak using agarose gel electrophoresis and flow cytometric analysis, respectively. The UVB-induced poly (ADP-ribose) polymerase-1 (PARP-1) cleavage, caspase activation and Bax/Bcl2 ratio were modulated following NG treatment, indicating an antiapoptotic effect of NG in UVB-damaged cells that occurs at least in part via caspase cascade pathway. Moreover, treatment of UVB-irradiated HaCaT cells with NG enhances the removal of CPD from the genome, as observed by both direct quantitation of CPD in genomic DNA and immuno-localization of the damage within the nuclei. The study provides a molecular basis for the action of NG as a promising natural flavonoid in preventing skin aging and carcinogenesis.
Many naturally occurring agents are believed to protect against UV-induced skin damage. In this study, we have investigated the effects of naringenin (NG), a naturally occurring citrus flavonone, on the removal of UVB-induced cyclobutane pyrimidine dimers (CPD) from the genome and apoptosis in immortalized p53-mutant human keratinocyte HaCaT cells. The colony-forming assay shows that treatment with NG significantly increases long-term cell survival after UVB irradiation. NG treatment also protects the cells from UVB-induced apoptosis, as indicated by the absence of the 180 base pair DNA ladders and the appearance of sub-G1 peak using agarose gel electrophoresis and flow cytometric analysis, respectively. The UVB-induced poly (ADP-ribose) polymerase-1 (PARP-1) cleavage, caspase activation and Bax/Bcl2 ratio were modulated following NG treatment, indicating an antiapoptotic effect of NG in UVB-damaged cells that occurs at least in part via caspase cascade pathway. Moreover, treatment of UVB-irradiated HaCaT cells with NG enhances the removal of CPD from the genome, as observed by both direct quantitation of CPD in genomic DNA and immuno-localization of the damage within the nuclei. The study provides a molecular basis for the action of NG as a promising natural flavonoid in preventing skin aging and carcinogenesis.Many naturally occurring agents are believed to protect against UV-induced skin damage. In this study, we have investigated the effects of naringenin (NG), a naturally occurring citrus flavonone, on the removal of UVB-induced cyclobutane pyrimidine dimers (CPD) from the genome and apoptosis in immortalized p53-mutant human keratinocyte HaCaT cells. The colony-forming assay shows that treatment with NG significantly increases long-term cell survival after UVB irradiation. NG treatment also protects the cells from UVB-induced apoptosis, as indicated by the absence of the 180 base pair DNA ladders and the appearance of sub-G1 peak using agarose gel electrophoresis and flow cytometric analysis, respectively. The UVB-induced poly (ADP-ribose) polymerase-1 (PARP-1) cleavage, caspase activation and Bax/Bcl2 ratio were modulated following NG treatment, indicating an antiapoptotic effect of NG in UVB-damaged cells that occurs at least in part via caspase cascade pathway. Moreover, treatment of UVB-irradiated HaCaT cells with NG enhances the removal of CPD from the genome, as observed by both direct quantitation of CPD in genomic DNA and immuno-localization of the damage within the nuclei. The study provides a molecular basis for the action of NG as a promising natural flavonoid in preventing skin aging and carcinogenesis.
AbstractMany naturally occurring agents are believed to protect against UV-induced skin damage. In this study, we have investigated the effects of naringenin (NG), a naturally occurring citrus flavonone, on the removal of UVB-induced cyclobutane pyrimidine dimers (CPD) from the genome and apoptosis in immortalized p53-mutant human keratinocyte HaCaT cells. The colony-forming assay shows that treatment with NG significantly increases long-term cell survival after UVB irradiation. NG treatment also protects the cells from UVB-induced apoptosis, as indicated by the absence of the 180base pair DNA ladders and the appearance of sub-G1 peak using agarose gel electrophoresis and flow cytometric analysis, respectively. The UVB-induced poly (ADP-ribose) polymerase-1 (PARP-1) cleavage, caspase activation and Bax/Bcl2 ratio were modulated following NG treatment, indicating an antiapoptotic effect of NG in UVB-damaged cells that occurs at least in part via caspase cascade pathway. Moreover, treatment of UVB-irradiated HaCaT cells with NG enhances the removal of CPD from the genome, as observed by both direct quantitation of CPD in genomic DNA and immuno-localization of the damage within the nuclei. The study provides a molecular basis for the action of NG as a promising natural flavonoid in preventing skin aging and carcinogenesis.
Author Zhao, Qun
Wani, Altaf A.
Patnaik, Srinivas
Wang, Qi-En
El-Mahdy, Mohamed A.
Arafa, El-Shaimaa
Wani, Gulzar
Zhu, Qianzheng
Mir, Safita N.
Barakat, Bassant
AuthorAffiliation 3 James Cancer Hospital and Research Institute, The Ohio State University, Columbus, OH
1 Department of Radiology, The Ohio State University, Columbus, OH
2 Biochemistry Program, The Ohio State University, Columbus, OH
AuthorAffiliation_xml – name: 1 Department of Radiology, The Ohio State University, Columbus, OH
– name: 2 Biochemistry Program, The Ohio State University, Columbus, OH
– name: 3 James Cancer Hospital and Research Institute, The Ohio State University, Columbus, OH
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  givenname: Mohamed A.
  surname: El-Mahdy
  fullname: El-Mahdy, Mohamed A.
  organization: Department of Radiology, The Ohio State University, Columbus, OH
– sequence: 2
  givenname: Qianzheng
  surname: Zhu
  fullname: Zhu, Qianzheng
  organization: Department of Radiology, The Ohio State University, Columbus, OH
– sequence: 3
  givenname: Qi-En
  surname: Wang
  fullname: Wang, Qi-En
  organization: Department of Radiology, The Ohio State University, Columbus, OH
– sequence: 4
  givenname: Gulzar
  surname: Wani
  fullname: Wani, Gulzar
  organization: Department of Radiology, The Ohio State University, Columbus, OH
– sequence: 5
  givenname: Srinivas
  surname: Patnaik
  fullname: Patnaik, Srinivas
  organization: Department of Radiology, The Ohio State University, Columbus, OH
– sequence: 6
  givenname: Qun
  surname: Zhao
  fullname: Zhao, Qun
  organization: Department of Radiology, The Ohio State University, Columbus, OH
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  givenname: El-Shaimaa
  surname: Arafa
  fullname: Arafa, El-Shaimaa
  organization: Department of Radiology, The Ohio State University, Columbus, OH
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  surname: Barakat
  fullname: Barakat, Bassant
  organization: Department of Radiology, The Ohio State University, Columbus, OH
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  givenname: Safita N.
  surname: Mir
  fullname: Mir, Safita N.
  organization: Department of Radiology, The Ohio State University, Columbus, OH
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  givenname: Altaf A.
  surname: Wani
  fullname: Wani, Altaf A.
  email: Wani.2@osu.edu
  organization: Department of Radiology, The Ohio State University, Columbus, OH
BackLink https://www.ncbi.nlm.nih.gov/pubmed/18086244$$D View this record in MEDLINE/PubMed
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This paper is part of a special issue dedicated to Professor Hasan Mukhtar on the occasion of his 60th birthday.
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PublicationTitle Photochemistry and photobiology
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Publisher Blackwell Publishing Ltd
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Snippet Many naturally occurring agents are believed to protect against UV‐induced skin damage. In this study, we have investigated the effects of naringenin (NG), a...
Many naturally occurring agents are believed to protect against UV-induced skin damage. In this study, we have investigated the effects of naringenin (NG), a...
UV exposure also causes DNA damage including UV-induced CPD and 6-4PP and these adducts can be removed by nucleotide excision repair (NER). Monoclonal...
AbstractMany naturally occurring agents are believed to protect against UV-induced skin damage. In this study, we have investigated the effects of naringenin...
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pubmed
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SourceType Open Access Repository
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StartPage 307
SubjectTerms Amino Acid Sequence
Apoptosis
Apoptosis - drug effects
Apoptosis - radiation effects
Biotechnology
Cell Line
Chemicals
Citrus
Deoxyribonucleic acid
DNA
DNA repair
Flavanones - pharmacology
Free radicals
Genetic disorders
Genome, Human
Human subjects
Humans
Keratinocytes - drug effects
Keratinocytes - radiation effects
Molecular Sequence Data
Monoclonal antibodies
Mortality
Pyrimidine Dimers - isolation & purification
Signal transduction
Ultraviolet Rays
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Title Naringenin Protects HaCaT Human Keratinocytes Against UVB-induced Apoptosis and Enhances the Removal of Cyclobutane Pyrimidine Dimers from the Genome
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