Ponce R A, Kavanagh T J, Mottet N K, Whittaker S G, Faustman E M
Department of Environmental Health, University of Washington, Seattle 98195.
Toxicol Appl Pharmacol. 1994 Jul;127(1):83-90. doi: 10.1006/taap.1994.1142.
Methyl mercury (MeHg) may interfere with cell cycle progression in a number of ways, most notably through an inhibition of protein synthesis or through effects on mitotic spindle performance; both mechanisms have experimental support. Results from investigations into the effects of MeHg exposure on cell cycle progression in a primary fetal rat CNS culture are presented here. Colchicine was also investigated because it is a well-characterized mitotic inhibitor. Flow cytometric DNA content analysis was utilized to determine the cell cycle distribution histograms of control and treated cultures. In addition, a flow cytometric technique which involves the incorporation of 5-bromo-2'-deoxyuridine into newly synthesized DNA was used to discriminate between successive cell cycles. Exposure of the CNS cell cultures to MeHg (2 and 4 microM) over a period of 0-48 hr led to a G2/M-phase inhibition as determined by flow cytometric DNA content analysis. Whereas exposure to 2 microM MeHg resulted in G2/M-phase inhibition, an analysis of cell cycle progression demonstrated an inhibition of cell cycling through any phase following exposure to 4 microM MeHg; these effects occurred in the first round of cell division following plating. Exposure to colchicine (25 nM) resulted in a G2/M-phase arrest similar to that observed with MeHg (2 microM). However, a comparison of the cytotoxicity patterns between MeHg-treated and colchicine-treated cultures suggests that MeHg-induced cytotoxicity cannot be solely ascribed to G2/M-phase arrest, since at equivalent levels of G2/M-phase arrest, MeHg was more cytotoxic than colchicine. These results are consistent with the hypothesis that microtubules, and the mitotic spindle, are especially sensitive to MeHg exposure.
甲基汞(MeHg)可能通过多种方式干扰细胞周期进程,最显著的是通过抑制蛋白质合成或影响有丝分裂纺锤体功能;这两种机制都有实验依据。本文展示了对原代胎鼠中枢神经系统培养物中甲基汞暴露对细胞周期进程影响的研究结果。秋水仙碱也在研究范围内,因为它是一种特征明确的有丝分裂抑制剂。利用流式细胞术DNA含量分析来确定对照培养物和处理后培养物的细胞周期分布直方图。此外,一种涉及将5-溴-2'-脱氧尿苷掺入新合成DNA的流式细胞术技术被用于区分连续的细胞周期。中枢神经系统细胞培养物在0 - 48小时内暴露于甲基汞(2和4微摩尔),通过流式细胞术DNA含量分析确定导致了G2/M期抑制。虽然暴露于2微摩尔甲基汞导致G2/M期抑制,但细胞周期进程分析表明,暴露于4微摩尔甲基汞后,细胞在任何阶段的循环都受到抑制;这些影响发生在接种后的第一轮细胞分裂中。暴露于秋水仙碱(25纳摩尔)导致的G2/M期阻滞与甲基汞(2微摩尔)观察到的相似。然而,甲基汞处理组和秋水仙碱处理组培养物细胞毒性模式的比较表明,甲基汞诱导的细胞毒性不能仅仅归因于G2/M期阻滞,因为在同等水平的G2/M期阻滞下,甲基汞比秋水仙碱更具细胞毒性。这些结果与微管和有丝分裂纺锤体对甲基汞暴露特别敏感的假设一致。