Unsuitability of lymphoblastoid cell lines as surrogate of cryopreserved isolated lymphocytes for the analysis of DNA double-strand break repair activity.

2.50
Hdl Handle:
http://hdl.handle.net/10146/114967
Title:
Unsuitability of lymphoblastoid cell lines as surrogate of cryopreserved isolated lymphocytes for the analysis of DNA double-strand break repair activity.
Authors:
Zijno, Andrea; Porcedda, Paola; Saini, Francesca; Allione, Alessandra; Garofalo, Bruno; Marcon, Francesca; Guarrera, Simonetta; Turinetto, Valentina; Minieri, Valentina; Funaro, Ada; Crebelli, Riccardo; Giachino, Claudia; Matullo, Giuseppe
Abstract:
As first task of a comprehensive investigation on DNA repair genotype-phenotype correlations, the suitability of Epstein-Barr virus (EBV)-transformed lymphoblastoid cell lines (LCLs) as surrogate of cryopreserved peripheral blood mononuclear cells (PBMCs) in DNA repair phenotypic assays was evaluated. To this aim the amount of DNA damage induced by gamma-rays and DNA repair capacity were evaluated in unstimulated (G(0)) and mitogen-simulated (G(2)) PBMC from 20 healthy subjects and in EBV-transformed LCL obtained from the same individuals. Phosphorylation of histone H2AX, micronuclei and chromosomal aberrations were the end-points investigated. The results obtained show higher basal frequencies of binucleated cells bearing micronuclei and nucleoplasmic bridge (NPB) in LCL with respect to PBMC, suggesting that EBV transformation may be associated with chromosomal instability. After irradiation, higher levels of micronuclei were induced in G(0)-treated PBMC compared to cycling LCL; conversely, NPB were more frequent in LCL than in PBMC. Moreover, higher levels of chromosomal aberrations were observed in G(2)-treated PBMC compared to LCL. Concerning gamma-H2AX measurements, phosphorylation levels 1h after treatment and dephosphorylation kinetics were basically similar in LCL and in PBMC. However, while Spearman's test showed a strong correlation between the results obtained in replicated experiments with PBMC, high inter-experimental variability and poor reproducibility was observed in the experiments performed with LCL, possibly due to the intrinsic instability of LCL. In summary, both the analysis of gamma-H2AX and the evaluation of chromosome damage highlighted a larger inter-experimental variability in the results obtained with LCL compared to PBMC. Noteworthy, the two set of results proved to lack any significant correlation at the individual level. These results indicate that LCL may be unsuitable for investigating genotype-phenotype correlations with phenotypic DNA repair assays, especially when low impact functional genetic variants are involved.
Citation:
Mutat. Res. 2010, 684 (1-2):98-105
Journal:
Mutation Research
Issue Date:
3-Feb-2010
URI:
http://hdl.handle.net/10146/114967
DOI:
10.1016/j.mrfmmm.2009.12.008
PubMed ID:
20035771
Additional Links:
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T2C-4Y0T99J-1&_user=1843694&_coverDate=02%2F03%2F2010&_rdoc=1&_fmt=high&_orig=search&_origin=search&_sort=d&_docanchor=&view=c&_acct=C000055040&_version=1&_urlVersion=0&_userid=1843694&md5=33eb0b69a15b8466f34140ad4bbe0934&searchtype=a
Type:
Article
Language:
en
ISSN:
0027-5107
Sponsors:
This work was supported by a grant of the Associazione Italiana per le Ricerche sul Cancro (Italy; G.M., A.Z., C.G.), of the Environmental Cancer Risk Nutrition and Individual Susceptibility project (G.M.), a network of excellence operating within the European Union Sixth Framework Program, Priority 5: ‘Food Quality and Safety’ (Contract No. 513943) and of the Agenzia Spaziale Italiana (A.Z.). A grant contribution has also been provided by the Programma Integrato Oncologia (PIO) (G.M.) and by Regione Piemonte Ricerca Sanitaria Finalizzata 2008 and 2009 (G.M. and C.G.). A grant contribution has been given by the Progetto Ricerca Sanitaria Finalizzata Regione Piemonte 2008 (A.F.).
Appears in Collections:
Articles

Full metadata record

DC FieldValue Language
dc.contributor.authorZijno, Andreaen
dc.contributor.authorPorcedda, Paolaen
dc.contributor.authorSaini, Francescaen
dc.contributor.authorAllione, Alessandraen
dc.contributor.authorGarofalo, Brunoen
dc.contributor.authorMarcon, Francescaen
dc.contributor.authorGuarrera, Simonettaen
dc.contributor.authorTurinetto, Valentinaen
dc.contributor.authorMinieri, Valentinaen
dc.contributor.authorFunaro, Adaen
dc.contributor.authorCrebelli, Riccardoen
dc.contributor.authorGiachino, Claudiaen
dc.contributor.authorMatullo, Giuseppeen
dc.date.accessioned2010-11-08T12:14:31Z-
dc.date.available2010-11-08T12:14:31Z-
dc.date.issued2010-02-03-
dc.identifier.citationMutat. Res. 2010, 684 (1-2):98-105en
dc.identifier.issn0027-5107-
dc.identifier.pmid20035771-
dc.identifier.doi10.1016/j.mrfmmm.2009.12.008-
dc.identifier.urihttp://hdl.handle.net/10146/114967-
dc.description.abstractAs first task of a comprehensive investigation on DNA repair genotype-phenotype correlations, the suitability of Epstein-Barr virus (EBV)-transformed lymphoblastoid cell lines (LCLs) as surrogate of cryopreserved peripheral blood mononuclear cells (PBMCs) in DNA repair phenotypic assays was evaluated. To this aim the amount of DNA damage induced by gamma-rays and DNA repair capacity were evaluated in unstimulated (G(0)) and mitogen-simulated (G(2)) PBMC from 20 healthy subjects and in EBV-transformed LCL obtained from the same individuals. Phosphorylation of histone H2AX, micronuclei and chromosomal aberrations were the end-points investigated. The results obtained show higher basal frequencies of binucleated cells bearing micronuclei and nucleoplasmic bridge (NPB) in LCL with respect to PBMC, suggesting that EBV transformation may be associated with chromosomal instability. After irradiation, higher levels of micronuclei were induced in G(0)-treated PBMC compared to cycling LCL; conversely, NPB were more frequent in LCL than in PBMC. Moreover, higher levels of chromosomal aberrations were observed in G(2)-treated PBMC compared to LCL. Concerning gamma-H2AX measurements, phosphorylation levels 1h after treatment and dephosphorylation kinetics were basically similar in LCL and in PBMC. However, while Spearman's test showed a strong correlation between the results obtained in replicated experiments with PBMC, high inter-experimental variability and poor reproducibility was observed in the experiments performed with LCL, possibly due to the intrinsic instability of LCL. In summary, both the analysis of gamma-H2AX and the evaluation of chromosome damage highlighted a larger inter-experimental variability in the results obtained with LCL compared to PBMC. Noteworthy, the two set of results proved to lack any significant correlation at the individual level. These results indicate that LCL may be unsuitable for investigating genotype-phenotype correlations with phenotypic DNA repair assays, especially when low impact functional genetic variants are involved.en
dc.description.sponsorshipThis work was supported by a grant of the Associazione Italiana per le Ricerche sul Cancro (Italy; G.M., A.Z., C.G.), of the Environmental Cancer Risk Nutrition and Individual Susceptibility project (G.M.), a network of excellence operating within the European Union Sixth Framework Program, Priority 5: ‘Food Quality and Safety’ (Contract No. 513943) and of the Agenzia Spaziale Italiana (A.Z.). A grant contribution has also been provided by the Programma Integrato Oncologia (PIO) (G.M.) and by Regione Piemonte Ricerca Sanitaria Finalizzata 2008 and 2009 (G.M. and C.G.). A grant contribution has been given by the Progetto Ricerca Sanitaria Finalizzata Regione Piemonte 2008 (A.F.).en
dc.language.isoenen
dc.relation.urlhttp://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T2C-4Y0T99J-1&_user=1843694&_coverDate=02%2F03%2F2010&_rdoc=1&_fmt=high&_orig=search&_origin=search&_sort=d&_docanchor=&view=c&_acct=C000055040&_version=1&_urlVersion=0&_userid=1843694&md5=33eb0b69a15b8466f34140ad4bbe0934&searchtype=aen
dc.subjectChromosome Aberrationsen
dc.subjectDNA Breaks, Double-Strandeden
dc.subjectDNA Repairen
dc.subjectCell Line, Transformeden
dc.subjectCell Transformation, Viralen
dc.subjectLeukocytes, Mononuclearen
dc.subjectLymphocytes/radiation effectsen
dc.subjectFlow Cytometryen
dc.subject.meshCell Line-
dc.subject.meshCell Line, Transformed-
dc.subject.meshCell Transformation, Viral-
dc.subject.meshChromosome Aberrations-
dc.subject.meshDNA Breaks, Double-Stranded-
dc.subject.meshDNA Repair-
dc.subject.meshFlow Cytometry-
dc.subject.meshG2 Phase-
dc.subject.meshGenetic Association Studies-
dc.subject.meshHistones-
dc.subject.meshLeukocytes, Mononuclear-
dc.subject.meshLymphocytes-
dc.subject.meshPhosphorylation-
dc.titleUnsuitability of lymphoblastoid cell lines as surrogate of cryopreserved isolated lymphocytes for the analysis of DNA double-strand break repair activity.en
dc.typeArticleen
dc.identifier.journalMutation Researchen

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