Inhibition by vitamin C of apoptosis induced by N-nitrosamines in HepG2 and HL-60 cells.

2.50
Hdl Handle:
http://hdl.handle.net/10146/76195
Title:
Inhibition by vitamin C of apoptosis induced by N-nitrosamines in HepG2 and HL-60 cells.
Authors:
Arranz, Nuria; Haza, Ana I.; Garcia, Almudena; Delgado, Ma Eugenia; Rafter, Joseph; Morales, Paloma
Abstract:
The aim of this study was to evaluate the effect of vitamin C towards N-nitrosopyrrolidine (NPYR)- and N-nitrosodimethylamine (NDMA)-induced apoptosis in human hepatoma (HepG2) and leukemia (HL-60) cell lines using flow cytometry analysis and the terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling assay (TUNEL). None of the vitamin C concentrations tested (1-100 microM) caused cytotoxicity in HepG2 cells. However, there were significant losses of HL-60 cells viability, measured by MTT assay, 72 h after treatment with 50 and 100 microM vitamin C (29 and 46%, respectively). Moreover, an increase of lactate dehydrogenase release was significant with 50 microM at 72 h (28%) and with 100 microM of vitamin C at 48 and 72 h (27 and 36%, respectively). Also, the percentage of apoptotic HL-60 cells found in TUNEL assay increased to 21% when they were treated with 100 microM vitamin C for 72 h. Thus, in subsequent simultaneous treatments with NPYR (30 and 50 mM) or NDMA (27 and 68 mM) and vitamin C, concentrations of 5-50 microM vitamin C were used. Our results revealed that vitamin C, at all concentrations and times tested, reduced the apoptosis induced by NPYR and NDMA in both cell lines, showing a similar effect in HepG2 and HL-60 cells towards NPYR (50 mM)--65 and 63% of reduction, respectively--whereas towards NDMA (27 mM) the inhibition was higher in HL-60 than in HepG2 cells--75 and 57%, respectively. Therefore, our findings suggest that inhibition of apoptosis may be one of the mechanisms by which vitamin C exerts its protective effect.
Citation:
J. Appl. Toxicol. 2008, 28 (6):788-796
Journal:
Journal of Applied Toxicology : JAT
Issue Date:
Aug-2008
URI:
http://hdl.handle.net/10146/76195
DOI:
10.1002/jat.1340
PubMed ID:
18344201
Additional Links:
http://www3.interscience.wiley.com/journal/117935040/abstract
Type:
Article
Language:
en
ISSN:
0260-437X
Appears in Collections:
Articles

Full metadata record

DC FieldValue Language
dc.contributor.authorArranz, Nuria-
dc.contributor.authorHaza, Ana I.-
dc.contributor.authorGarcia, Almudena-
dc.contributor.authorDelgado, Ma Eugenia-
dc.contributor.authorRafter, Joseph-
dc.contributor.authorMorales, Paloma-
dc.date.accessioned2009-08-04T08:20:14Z-
dc.date.available2009-08-04T08:20:14Z-
dc.date.issued2008-08-
dc.identifier.citationJ. Appl. Toxicol. 2008, 28 (6):788-796en
dc.identifier.issn0260-437X-
dc.identifier.pmid18344201-
dc.identifier.doi10.1002/jat.1340-
dc.identifier.urihttp://hdl.handle.net/10146/76195-
dc.description.abstractThe aim of this study was to evaluate the effect of vitamin C towards N-nitrosopyrrolidine (NPYR)- and N-nitrosodimethylamine (NDMA)-induced apoptosis in human hepatoma (HepG2) and leukemia (HL-60) cell lines using flow cytometry analysis and the terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling assay (TUNEL). None of the vitamin C concentrations tested (1-100 microM) caused cytotoxicity in HepG2 cells. However, there were significant losses of HL-60 cells viability, measured by MTT assay, 72 h after treatment with 50 and 100 microM vitamin C (29 and 46%, respectively). Moreover, an increase of lactate dehydrogenase release was significant with 50 microM at 72 h (28%) and with 100 microM of vitamin C at 48 and 72 h (27 and 36%, respectively). Also, the percentage of apoptotic HL-60 cells found in TUNEL assay increased to 21% when they were treated with 100 microM vitamin C for 72 h. Thus, in subsequent simultaneous treatments with NPYR (30 and 50 mM) or NDMA (27 and 68 mM) and vitamin C, concentrations of 5-50 microM vitamin C were used. Our results revealed that vitamin C, at all concentrations and times tested, reduced the apoptosis induced by NPYR and NDMA in both cell lines, showing a similar effect in HepG2 and HL-60 cells towards NPYR (50 mM)--65 and 63% of reduction, respectively--whereas towards NDMA (27 mM) the inhibition was higher in HL-60 than in HepG2 cells--75 and 57%, respectively. Therefore, our findings suggest that inhibition of apoptosis may be one of the mechanisms by which vitamin C exerts its protective effect.en
dc.language.isoenen
dc.relation.urlhttp://www3.interscience.wiley.com/journal/117935040/abstracten
dc.subjectNPYRen
dc.subjectNDMAen
dc.subjectvitamin Cen
dc.subjectHepG2 cellsen
dc.subjectHL-60 cellsen
dc.subjectapoptosisen
dc.subject.meshAntioxidants-
dc.subject.meshApoptosis-
dc.subject.meshAscorbic Acid-
dc.subject.meshCarcinogens-
dc.subject.meshCell Line, Tumor-
dc.subject.meshCell Survival-
dc.subject.meshCytochrome P-450 Enzyme System-
dc.subject.meshExcitatory Amino Acid Agonists-
dc.subject.meshHL-60 Cells-
dc.subject.meshHumans-
dc.subject.meshIn Situ Nick-End Labeling-
dc.subject.meshL-Lactate Dehydrogenase-
dc.subject.meshLiver Neoplasms-
dc.subject.meshN-Methylaspartate-
dc.subject.meshN-Nitrosopyrrolidine-
dc.subject.meshNitrosamines-
dc.subject.meshTetrazolium Salts-
dc.subject.meshThiazoles-
dc.titleInhibition by vitamin C of apoptosis induced by N-nitrosamines in HepG2 and HL-60 cells.en
dc.typeArticleen
dc.identifier.journalJournal of Applied Toxicology : JATen

Related articles on PubMed

All Items in ECNIS-NIOM are protected by copyright, with all rights reserved, unless otherwise indicated.