Mukherjee A B, Costello C
Department of Biological Sciences, Fordham University, Bronx, NY 10458, USA.
Mech Ageing Dev. 1998 Jun 15;103(2):209-22. doi: 10.1016/s0047-6374(98)00041-4.
Cultured cells from human premature aging syndromes show premature replicative senescence and as such might serve as models for genetic studies of cellular replicative senescence/aging. To date, no systematic study on chromosome-specific aneuploidy in premature aging syndromes has been carried out using molecular-cytogenetic techniques. We, therefore, have performed a comparative analysis of chromosome-specific aneuploidy levels at both interphase and metaphase in earlier (younger) and later (older) passage fibroblasts from two normal male and female versus six male and female premature aging syndromes (Cockayne, Hutchinson-Gilford and Werner) by FISH. We have used seven chromosome-specific DNA probes (nos. 1, 4, 6, 8, 10, 15, X) and four DNA probes (nos. 1, 4, 6, X) for females and males, respectively. Our data on total aneuploidy of each chromosome in all cell cultures indicated that significantly higher percentages of aneuploid cells were detectable at interphase than at metaphase. Also, the interphase aneuploidy levels of all chromosomes under study were significantly higher in cells from the syndromes as compared to these of the normal controls at both earlier and later passages. In general, the interphase aneuploidy level of each of the chromosomes in both the control and experimental cell cultures increased with in vitro proliferation and aging, although to a much lesser extent in the controls. The aneuploidy levels, however, varied widely from chromosome to chromosome in each case. Since adequate numbers of mitotic cells were not always available in some later passage cells of various premature aging syndromes, metaphase chromosome analysis for all chromosomes under study was not always possible at this stage, but interphase cytogenetics was very informative in all cases. No consistent pattern of chromosome-specific aneuploidy was detected in the earlier passage cells (younger cells) of the syndromes. However, the later passage cells (older cells) from all three female syndromes consistently showed the highest aneuploidy levels for the X chromosome at interphase.
来自人类早衰综合征的培养细胞表现出过早的复制性衰老，因此可作为细胞复制性衰老/老化遗传研究的模型。迄今为止，尚未使用分子细胞遗传学技术对早衰综合征中染色体特异性非整倍体进行系统研究。因此，我们通过荧光原位杂交（FISH）对来自两名正常男性和女性以及六种男性和女性早衰综合征（科凯恩综合征、哈钦森-吉尔福德综合征和沃纳综合征）的早期（较年轻）和晚期（较老）传代成纤维细胞在间期和中期的染色体特异性非整倍体水平进行了比较分析。我们分别使用了七种染色体特异性DNA探针（1、4、6、8、10、15、X号染色体）以及针对女性和男性的四种DNA探针（1、4、6、X号染色体）。我们关于所有细胞培养物中每条染色体总非整倍体的数据表明，间期可检测到的非整倍体细胞百分比显著高于中期。此外，在早期和晚期传代时，与正常对照相比，综合征细胞中所有研究染色体的间期非整倍体水平均显著更高。总体而言，对照和实验细胞培养物中每条染色体的间期非整倍体水平均随体外增殖和老化而增加，尽管在对照中增加幅度小得多。然而，每种情况下各染色体的非整倍体水平差异很大。由于在各种早衰综合征的一些晚期传代细胞中并非总能获得足够数量的有丝分裂细胞，因此现阶段并非总能对所有研究染色体进行中期染色体分析，但间期细胞遗传学在所有情况下都提供了非常丰富的信息。在综合征的早期传代细胞（较年轻细胞）中未检测到一致的染色体特异性非整倍体模式。然而，来自所有三种女性综合征的晚期传代细胞（较老细胞）在间期始终显示出X染色体的非整倍体水平最高。
