Variable NAT1 enzyme activity in long-term cultured human HaCaT keratinocytes.
Identifieur interne : 000487 ( PubMed/Corpus ); précédent : 000486; suivant : 000488Variable NAT1 enzyme activity in long-term cultured human HaCaT keratinocytes.
Auteurs : Simone Scheitza ; Jutta Bonifas ; Brunhilde BlömekeSource :
- Journal of toxicology and environmental health. Part A [ 1528-7394 ] ; 2012.
English descriptors
- KwdEn :
- Arylamine N-Acetyltransferase (metabolism), Cell Division (physiology), Cell Line, Cell Proliferation, Cells, Cultured, Cytological Techniques, Freezing, Humans, Isoenzymes (metabolism), Keratin-1 (metabolism), Keratinocytes (enzymology), Real-Time Polymerase Chain Reaction, Transglutaminases (metabolism).
- MESH :
- chemical , metabolism : Arylamine N-Acetyltransferase, Isoenzymes, Keratin-1, Transglutaminases.
- enzymology : Keratinocytes.
- physiology : Cell Division.
- Cell Line, Cell Proliferation, Cells, Cultured, Cytological Techniques, Freezing, Humans, Real-Time Polymerase Chain Reaction.
Abstract
Since animal testing should be avoided whenever possible, the development of in vitro tests for predicting the effect of chemicals becomes a major field. This rise of in vitro test systems led to an increased requirement for well-characterized continuously growing cell lines. Monitoring of the cells during test and routine culture is necessary to gain relevant and reproducible results. In the present study, the influence of passaging under constant culture conditions on the human keratinocyte cell line HaCaT was investigated. Data demonstrated that growth rate rose with increasing passages. Doubling times of the cells were decreased to 24 ± 0.6 h in the late passages (12-16), in comparison to 36.2 ± 1.5 h in the early passages (2-8). These data were confirmed by a fall in mRNA expression levels of keratin 1 and transglutaminase 1 within the passages. Furthermore, the activities of the xenobiotic metabolizing phase II enzyme N-acetyltransferase 1 (NAT1) were higher in the late passages compared to the early passages. These results are contrary to an expected decrease in enzyme activity and proliferation rate induced by replicative senescence or cell aging. Data also indicate that routine culture might result in significant changes in proliferation and phase II metabolism. These findings reinforce the necessity of a strict characterization and knowledge of regulation of in vitro systems, as well as the need for new biomarkers, in order to use cells for the development and evaluation of reproducible in vitro test systems.
DOI: 10.1080/15287394.2012.674915
PubMed: 22686306
Links to Exploration step
pubmed:22686306Le document en format XML
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This rise of in vitro test systems led to an increased requirement for well-characterized continuously growing cell lines. Monitoring of the cells during test and routine culture is necessary to gain relevant and reproducible results. In the present study, the influence of passaging under constant culture conditions on the human keratinocyte cell line HaCaT was investigated. Data demonstrated that growth rate rose with increasing passages. Doubling times of the cells were decreased to 24 ± 0.6 h in the late passages (12-16), in comparison to 36.2 ± 1.5 h in the early passages (2-8). These data were confirmed by a fall in mRNA expression levels of keratin 1 and transglutaminase 1 within the passages. Furthermore, the activities of the xenobiotic metabolizing phase II enzyme N-acetyltransferase 1 (NAT1) were higher in the late passages compared to the early passages. These results are contrary to an expected decrease in enzyme activity and proliferation rate induced by replicative senescence or cell aging. Data also indicate that routine culture might result in significant changes in proliferation and phase II metabolism. These findings reinforce the necessity of a strict characterization and knowledge of regulation of in vitro systems, as well as the need for new biomarkers, in order to use cells for the development and evaluation of reproducible in vitro test systems.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">22686306</PMID><DateCreated><Year>2012</Year><Month>06</Month><Day>12</Day></DateCreated><DateCompleted><Year>2012</Year><Month>08</Month><Day>16</Day></DateCompleted><DateRevised><Year>2012</Year><Month>06</Month><Day>12</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">1528-7394</ISSN><JournalIssue CitedMedium="Internet"><Volume>75</Volume><Issue>8-10</Issue><PubDate><Year>2012</Year></PubDate></JournalIssue><Title>Journal of toxicology and environmental health. Part A</Title><ISOAbbreviation>J. Toxicol. Environ. Health Part A</ISOAbbreviation></Journal><ArticleTitle>Variable NAT1 enzyme activity in long-term cultured human HaCaT keratinocytes.</ArticleTitle><Pagination><MedlinePgn>471-7</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1080/15287394.2012.674915</ELocationID><Abstract><AbstractText>Since animal testing should be avoided whenever possible, the development of in vitro tests for predicting the effect of chemicals becomes a major field. This rise of in vitro test systems led to an increased requirement for well-characterized continuously growing cell lines. Monitoring of the cells during test and routine culture is necessary to gain relevant and reproducible results. In the present study, the influence of passaging under constant culture conditions on the human keratinocyte cell line HaCaT was investigated. Data demonstrated that growth rate rose with increasing passages. Doubling times of the cells were decreased to 24 ± 0.6 h in the late passages (12-16), in comparison to 36.2 ± 1.5 h in the early passages (2-8). These data were confirmed by a fall in mRNA expression levels of keratin 1 and transglutaminase 1 within the passages. Furthermore, the activities of the xenobiotic metabolizing phase II enzyme N-acetyltransferase 1 (NAT1) were higher in the late passages compared to the early passages. These results are contrary to an expected decrease in enzyme activity and proliferation rate induced by replicative senescence or cell aging. Data also indicate that routine culture might result in significant changes in proliferation and phase II metabolism. These findings reinforce the necessity of a strict characterization and knowledge of regulation of in vitro systems, as well as the need for new biomarkers, in order to use cells for the development and evaluation of reproducible in vitro test systems.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Scheitza</LastName><ForeName>Simone</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>Department of Environmental Toxicology, University of Trier, Trier, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Bonifas</LastName><ForeName>Jutta</ForeName><Initials>J</Initials></Author><Author ValidYN="Y"><LastName>Blömeke</LastName><ForeName>Brunhilde</ForeName><Initials>B</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>J Toxicol Environ Health A</MedlineTA><NlmUniqueID>100960995</NlmUniqueID><ISSNLinking>0098-4108</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D007527">Isoenzymes</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D053556">Keratin-1</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 2.3.1.5</RegistryNumber><NameOfSubstance UI="D001191">Arylamine N-Acetyltransferase</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 2.3.1.5</RegistryNumber><NameOfSubstance UI="C089384">N-acetyltransferase 1</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 2.3.2.13</RegistryNumber><NameOfSubstance UI="D011503">Transglutaminases</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 2.3.2.13</RegistryNumber><NameOfSubstance UI="C105804">transglutaminase 1</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D001191" MajorTopicYN="N">Arylamine N-Acetyltransferase</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D002455" MajorTopicYN="N">Cell Division</DescriptorName><QualifierName UI="Q000502" MajorTopicYN="N">physiology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D002460" MajorTopicYN="N">Cell Line</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D049109" MajorTopicYN="N">Cell Proliferation</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002478" MajorTopicYN="N">Cells, Cultured</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D003584" MajorTopicYN="N">Cytological Techniques</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D005615" MajorTopicYN="N">Freezing</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D007527" MajorTopicYN="N">Isoenzymes</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D053556" MajorTopicYN="N">Keratin-1</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D015603" MajorTopicYN="N">Keratinocytes</DescriptorName><QualifierName UI="Q000201" MajorTopicYN="Y">enzymology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D060888" MajorTopicYN="N">Real-Time Polymerase Chain Reaction</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D011503" MajorTopicYN="N">Transglutaminases</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="entrez"><Year>2012</Year><Month>6</Month><Day>13</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2012</Year><Month>6</Month><Day>13</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2012</Year><Month>8</Month><Day>17</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">22686306</ArticleId><ArticleId IdType="doi">10.1080/15287394.2012.674915</ArticleId></ArticleIdList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
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