Marshall R R, Murphy M, Kirkland D J, Bentley K S
Corning Hazleton Europe, Harrogate, UK.
Mutat Res. 1996 Dec;372(2):233-45. doi: 10.1016/s0027-5107(96)00143-1.
Cytochalasin B-blocked binucleate human lymphocytes from female donors have been used to measure micronucleus induction and other aneuploidy events after treatment with colchicine, vinblastine or carbendazim. For the aneuploidy events, centromeric probes for 6 selected chromosomes (1, 8, X, 11, 17, 18) were used to measure chromosome loss, addition and non-disjunction in the interphase nuclei of these binucleate cells. The chromosomes were probed in pairs using Cy-3 (red) and FITC (green) labels for the 2 different centromeric regions. For colchicine, the total non-disjunction frequencies for chromosomes 1 and 8 were similar to the total micronucleus frequencies, but were detected as significant at lower concentrations. For vinblastine (chromosomes 1 and 8) and carbendazim (all 6 chromosomes) the frequencies of non-disjunction far exceeded (7 and > 2-fold, respectively) the peak frequencies of micronucleus induction. Although most chromosomes exhibited similar sensitivity in all the aneuploidy events measured, there was an indication that chromosome X was more than susceptible to non-disjunction than the other chromosomes. We believe that measurement of non-disjunction in binucleate human lymphocytes using chromosome specific centromeric probes offers a sensitive method for detection of aneuploidy and is particularly appropriate for the establishment of thresholds.
来自女性供体的细胞松弛素B阻断的双核人淋巴细胞已被用于测量秋水仙碱、长春碱或多菌灵处理后的微核诱导和其他非整倍体事件。对于非整倍体事件，使用针对6条选定染色体（1、8、X、11、17、18）的着丝粒探针来测量这些双核细胞间期核中的染色体丢失、增加和不分离情况。使用Cy-3（红色）和FITC（绿色）标签对2个不同的着丝粒区域进行染色体配对探测。对于秋水仙碱，染色体1和8的总不分离频率与总微核频率相似，但在较低浓度下被检测为显著。对于长春碱（染色体1和8）和多菌灵（所有6条染色体），不分离频率远远超过（分别为7倍和>2倍）微核诱导的峰值频率。尽管在所有测量的非整倍体事件中，大多数染色体表现出相似的敏感性，但有迹象表明，X染色体比其他染色体更容易发生不分离。我们认为，使用染色体特异性着丝粒探针测量双核人淋巴细胞中的不分离情况，为检测非整倍体提供了一种灵敏的方法，尤其适用于阈值的确定。
