Naslov
Application of cytogenetic endpoints and Comet assay on human lymphocytes treated with vincristine in vitro

Autori
Kopjar, Nevenka ; Garaj-Vrhovac, Vera

Izvornik
Neoplasma (0028-2685) 47 (2000), 3; 162-167

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
human lymphocytes; vincristine; in vitro; comet assay; cytogenetic endpoints

Sažetak
The genotoxic potential of vincristine is assessed on human peripheral blood lymphocytes following administration of the drug at dose 0.0875 ƒÝg/ml by use of analysis of structural chromosome aberrations (CA), micronucleus assay (MN), sister chromatid exchange (SCE) analysis and single cell gel electrophoresis - Comet assay (SCGE). In vitro treatment of human lymphocytes with vincristine was performed on cells in G0 phase as well on lymphocyte cultures 24 hours after stimulation with mitogen phytohaemagglutinine. For the Comet assay 24h, 48h and 72h after treatment treated cells were embedded in agarose on slides, lysed with alkaline lysis solution and exposed to an electric field. DNA migrated within the agarose and formed comets whose length depend on the amount of DNA damage. For the analysis of structural CA cells were grown on F-10 medium for 48 hours, and for MN and SCE analysis for 72 hours. The results on SCGE showed an increase in tail length compared to control both in cells treated in G0 and in cells treated 24 h after mitogen stimulation. The amount of DNA damage was higher in cells treated with vincristine 24 h after mitogen stimulation. Administered concentration of drug caused total inhibition of lymphocyte growth in 72 h cultures for MN and SCE analysis indicating strong microtubule distruptive effects of vincristine. Analysis of structural CA reveal chromatid breaks and acentric fragments as the main aberration types both in cells treated in G0 and in cells treated 24 h after mitogen stimulation. Number of these aberrations was higher in cells treated in G0 phase. Results obtained in this study by use of different cytogenetic endpoints confirmed that vincristine exhibit both aneugenic and clastogenic effects on human lymphocytes.

Izvorni jezik
Engleski

Znanstvena područja
Javno zdravstvo i zdravstvena zaštita

POVEZANOST RADA