IMPAIRED S-PHASE TRANSIT OF WERNER SYNDROME CELLS EXPRESSED IN LYMPHOBLASTOID CELL-LINES
IMPAIRED S-PHASE TRANSIT OF WERNER SYNDROME CELLS EXPRESSED IN LYMPHOBLASTOID CELL-LINES
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DOI：
10.1016/0014-4827(92)90074-i
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发表时间：
1992-10-01
影响因子：
3.7
通讯作者：
MARTIN, GM
中科院分区：
文献类型：
--
作者：
POOT, M;HOEHN, H;MARTIN, GM
沃纳综合征（WS）的临床表型包括身材矮小、早发性白内障、皮肤萎缩、骨质疏松、头发变白和脱发、肿瘤、糖尿病和动脉硬化。来自这种常染色体隐性遗传病患者的培养细胞表现出染色体不稳定性和显著降低的复制寿命和生长速度。为了阐明与生长缺陷相关的细胞周期改变，我们用5-溴脱氧尿嘧啶连续标记了来自5名WS患者和4名健康成人对照的淋巴样细胞系。双变量Hoechst 33258/溴化乙啶流式细胞术显示WS细胞S期最短持续时间延长2.4 h （P< 0.005）。此外，WS患者在S期不可逆阻滞的增殖细胞比例（5.4%比对照组的1.4%）显著升高（P< 0.001）。其他细胞周期区室在WS细胞系中未受显著影响。作为对WS表型是由于DNA拓扑异构酶I （topo I）或DNA拓扑异构酶II （topo II）缺陷的假设的部分检验，我们将健康对照的淋巴样细胞暴露于拓扑异构酶I抑制剂喜树碱或拓扑异构酶II抑制剂4 ' -（9-吖啶胺）甲磺酸-间茴香胺。这些化合物引起的细胞动力学改变不同于未经治疗的WS患者。因此，在WS中不太可能出现拓扑I或拓扑II的原发性缺陷。然而，我们的细胞周期结果提供了重要的证据，证明生化遗传病变实际上是在淋巴母细胞系中表达的，淋巴母细胞系是这些受试者中最容易获得的细胞。
The clinical phenotype of Werner's syndrome (WS) includes short stature, premature cataracts, skin atrophy, osteoporosis, graying and loss of hair, neoplasia, diabetes mellitus, and arteriosclerosis. Cultured cells from patients with this autosomal recessive disorder exhibit chromosomal instability and a markedly reduced replicative lifespan and growth rate. To elucidate the cell cycle alterations associated with the growth deficit, we continuously labeled lymphoid cell lines from five WS patients and from four healthy adult controls with 5-bromodeoxyuridine. Bivariate Hoechst 33258/ethidium bromide flow cytometry revealed a 2.4-h prolongation in the minimal duration of the S phase of WS cells (P< 0.005). Moreover, the fraction of proliferating cells irreversibly arrested in the S phase (5.4% vs 1.4% in controls) was significantly elevated in WS (P< 0.001). Other cell cycle compartments were not significantly affected in WS cell lines. As a partial test of the hypothesis that the WS phenotype is due to a defect in DNA topoisomerase I (topo I) or DNA topoisomerase II (topo II) we exposed lymphoid cells from a healthy control to the topo I inhibitor camptothecin or to the topo II inhibitor 4′-(9-acridinylamino)methanesulfon-m-anisidine. The cell kinetic alterations elicited by these compounds differed from that exhibited by untreated WS patients. Thus, a primary defect in topo I or II is unlikely in WS. Our cell cycle results, however, provide important evidence that the biochemical genetic lesion is in fact expressed in lymphoblastoid cell lines, the most readily available cells from such subjects.