Kopjar N, Garaj-Vrhovac V
Institute for Medical Research and Occupational Health, Zagreb, Croatia.
Neoplasma. 2000;47(3):162-7.
The genotoxic potential of vincristine is assessed on human peripheral blood lymphocytes following administration of the drug at a dose 0.0875 microg/ml by use of single cell gel electrophoresis - Comet assay (SCGE), analysis of structural chromosome aberrations (CA), micronucleus assay (MN) and sister chromatid exchange (SCE) analysis. 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 phytohemagglutinine. For the Comet assay at 24, 48 and 72 h the 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 depends 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 lymphocytes growth in 72-h cultures for MN and SCE analysis indicating strong microtubule distruptive effects of vincristine. Analysis of structural CA reveals 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 exhibits both aneugenic and clastogenic effects on human lymphocytes.
通过单细胞凝胶电泳 - 彗星试验（SCGE）、结构染色体畸变（CA）分析、微核试验（MN）和姐妹染色单体交换（SCE）分析，在以0.0875微克/毫升的剂量给予长春新碱后，评估其对人外周血淋巴细胞的遗传毒性潜力。用长春新碱对人淋巴细胞进行体外处理，处理对象为处于G0期的细胞以及在用丝裂原植物血凝素刺激24小时后的淋巴细胞培养物。对于在24、48和72小时进行的彗星试验，将处理后的细胞包埋在载玻片上的琼脂糖中，用碱性裂解液裂解并暴露于电场中。DNA在琼脂糖内迁移并形成彗星状，其长度取决于DNA损伤的程度。对于结构CA分析，细胞在F - 10培养基上生长48小时，对于MN和SCE分析则生长72小时。SCGE结果显示，与对照组相比，在G0期处理的细胞以及在丝裂原刺激后24小时处理的细胞中，尾长均增加。在丝裂原刺激后24小时用长春新碱处理的细胞中，DNA损伤量更高。在用于MN和SCE分析的72小时培养物中，所给药的药物浓度导致淋巴细胞生长完全抑制，表明长春新碱具有强烈的微管破坏作用。结构CA分析显示，在G0期处理的细胞以及在丝裂原刺激后24小时处理的细胞中，染色单体断裂和无着丝粒片段是主要的畸变类型。在G0期处理的细胞中，这些畸变的数量更高。本研究通过使用不同的细胞遗传学终点获得的结果证实，长春新碱对人淋巴细胞具有非整倍体诱导和染色体断裂效应。
