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[一种细胞外热休克蛋白对印度麂培养皮肤成纤维细胞染色体变异性的影响]

[Effect of an extracellular heat shock protein on chromosome variability in Indian muntjak cultured skin fibroblasts].

作者信息

Polianskaia G G, Kinev A V, Sakuta G A, Aseeva E V

机构信息

Institute of Cytology, Russian Academy of Sciences, St. Petersburg.

出版信息

Tsitologiia. 1997;39(11):1070-82.

PMID:9505350
Abstract

It is known that the essential function of extracellular HSP70 (e-HSP70) to protect cell processes, indirectly associated with the genetic structures. A direct influence of e-HSP70 on chromosomal stability has not been studied. This explains actuality of the suggested investigation. A study was made of the influence of e-HSP70 on chromosomal variability at different phases of the first mitotic cycle in both intact and ciprofloxacin (CF) treated cells of the Indian muntjac skin fibroblasts. E-HSP70 (10 mg/ml) exerts no influence on the level of chromosomal aberrations, typical of the control. After a joint action of e-HSP70 and CF (50 mg/ml) no influence was also exerted on the antibiotic induced genotoxicity effect. CF and e-HSP70 (50 and 100 mg/ml, resp.) acting apart on intact cells during 6 and 24 h induce a significant increase of chromosomal aberrations compared to the control, primarily at the expense of chromatid or chromosomal breaks (depending on the duration of respective effect). CF and e-HSP70 acting apart on intact cells during 6 h with the following cultivation for 18 h in the fresh medium prior to fixation also induce a significant increase of chromosomal aberrations compared to the control, primarily at the expense of both breaks and dicentrics (telomeric associations). The joint action of CF and e-HSP70 on cultivated cells during 6 and 24 h and when CF and e-HSP70 were added respectively on 24 and 6 h prior to fixation, a significant decrease in chromosomal aberrations compared to the control level was induced. A simultaneous addition of CF and e-HSP70 in 6 h with the following cultivation for 18 h in fresh medium prior to fixation exerted no influence on the degree of genotoxicity effect, typical of the separate action of these agents. However a significant decrease in the number of dicentrics occurred. Apparently, e-HSP70 has a protective effect on chromosomal stability mainly at phase 62 of the mitotic cycle. The denaturated e-HSP70 (50, 100 mg/ml) has a genotoxicity effect similar to that of the infact e-HSP70 under above conditions. The joint action of CF and denaturated e-HSP70 (50 mg/ml) during 24 h exerts no influence on the degree of genotoxicity effect, induced by the separate action of these agents or leads to an increase in the number of dicentrics, acting separately or jointly compared to the control. The denaturated e-HSP70 (100 mg/ml) acting jointly with CF for 24 h, increases the degree of genotoxicity effects, induced by separate actions of the agents. Lipopolysacharide (50 mg/ml) exerts no influence on the number of chromosomal aberrations in the control. The sensitivity of individual chromosomes and their regions to agents inducing chromosomal instability is different. The preferential involvement of some chromosomes in dicentric formation was observed. A possible role of dicentrics in adaptation of cells belonging to "markerless" lines to infavourable factors of the environment, and possible mechanisms of protecting effect exerted by e-HSP70 on chromosomes are discussed.

摘要

已知细胞外热休克蛋白70(e-HSP70)的基本功能是保护细胞过程,这与遗传结构间接相关。尚未研究e-HSP70对染色体稳定性的直接影响。这解释了所建议研究的现实意义。对印度麂皮成纤维细胞完整细胞和环丙沙星(CF)处理细胞的第一次有丝分裂周期不同阶段,e-HSP70对染色体变异性的影响进行了研究。e-HSP70(10毫克/毫升)对染色体畸变水平无影响,这是对照组的典型特征。e-HSP70和CF(50毫克/毫升)共同作用也未对抗生素诱导的遗传毒性效应产生影响。CF和e-HSP70(分别为50和100毫克/毫升)在6小时和24小时分别作用于完整细胞,与对照组相比,染色体畸变显著增加,主要是由于染色单体或染色体断裂(取决于各自作用的持续时间)。CF和e-HSP70在6小时分别作用于完整细胞,随后在固定前在新鲜培养基中培养18小时,与对照组相比,染色体畸变也显著增加,主要是由于断裂和双着丝粒(端粒关联)。CF和e-HSP70在6小时和24小时对培养细胞共同作用,以及在固定前分别在24小时和6小时添加CF和e-HSP70,与对照水平相比,染色体畸变显著降低。在6小时同时添加CF和e-HSP70,随后在固定前在新鲜培养基中培养18小时,对这些试剂单独作用典型的遗传毒性效应程度无影响。然而,双着丝粒数量显著减少。显然,e-HSP70主要在有丝分裂周期的62期对染色体稳定性具有保护作用。变性e-HSP70(50、100毫克/毫升)在上述条件下具有与天然e-HSP70相似的遗传毒性效应。CF和变性e-HSP70(50毫克/毫升)在24小时共同作用,对这些试剂单独作用诱导的遗传毒性效应程度无影响,或与对照组相比,单独或共同作用导致双着丝粒数量增加。变性e-HSP70(100毫克/毫升)与CF共同作用24小时,增加了试剂单独作用诱导的遗传毒性效应程度。脂多糖(50毫克/毫升)对对照组染色体畸变数量无影响。个别染色体及其区域对诱导染色体不稳定的试剂的敏感性不同。观察到一些染色体优先参与双着丝粒形成。讨论了双着丝粒在属于“无标记”系的细胞适应环境不利因素中的可能作用,以及e-HSP70对染色体发挥保护作用的可能机制。

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