Cell Biology and Toxicology, 2012, 28(5):331-342.

Aneugenic effects of the genistein glycosidic derivative substituted at C7 with the unsaturated disaccharide

Agnieszka Gogler-Pigłowska, Aleksandra Rusin, Daria Bochenek, et al

Center For Translational Research and Molecular Biology of CancerMaria Skłodowska-Curie Memorial Cancer Center and Institute of Oncology, Gliwice Branch, PolandGliwicePoland; and Department of Organic Chemistry, Bioorganic Chemistry and BiotechnologySilesian University of TechnologyGliwicePoland

Genistein, due to its recognized chemopreventive and antitumour potential, is a molecule of interest as a lead compound in drug design. Recently, we found that the novel genistein derivative, [7-O-(2,3,4,6-tetra-O-acetyl-β-D-galactopyranosyl)-(1 → 4)-(6-O-acetyl-hex-2-ene-α-D-erythro pyranosyl) genistein, named G21, induced aberrations in mitotic spindle formation. In the presented study, we investigated the properties of G21 relevant to its genotoxic activity. The inhibition of topoisomerase IIα activity was evaluated in decatenation assay and immunoband depletion assay, the covalent DNA-topoisomerase IIα complexes and histone ɣH2AX were detected immunofluorescently. Genotoxic effects of the tested compounds were assessed in micronucleation assay. The presence of centromeres in the micronuclei and the multiplication of centrosomes were evaluated in fluorescence immunolabelled specimens. The inhibition of tubulin polymerization was measured spectrophotometrically. We found that both tested drugs were able to inhibit topoisomerase II activity; however, G21, in contrast to genistein, blocked this enzyme at the concentration far exceeding cytotoxic IC(50). We also found that both compounds caused micronucleation in DU 145 prostate cancer cells, but in contrast to genistein, G21 exhibited aneugenic activity, manifested by the presence of centromeres in micronuclei formed in cells treated with the drug. Aneugenic properties of G21 resulted from the inhibition of tubulin polymerization and centrosome disruption, not observed in the presence of genistein. The study supports and extends our previous observations that the mechanisms of cytotoxicity of genistein and its new glycosidic derivative-G21 are significantly different.

荷兰《细胞生物学和毒理学》，2012年10月

金雀异黄素糖苷衍生物在 C7取代不饱和双糖的细胞遗传毒性效应

阿格涅斯卡 高格拉 皮格劳斯卡，亚历珊德拉 卢森，达瑞亚 保什尼克，等

波兰纪念玛利亚居里夫人癌症中心和肿瘤研究所转化研究与分子生物学中心；波兰西里西亚科技大学有机化学、 生物有机化学和生物技术学院

金雀异黄素，由于其公认的化学防癌和抗癌潜力，在药物设计中被视为一种先导化合物，是一种吸引人的分子。最近，我们发现，新型的金雀异黄素衍生物，[7-氧-(2,3,4,6-四-氧-乙酰基-β-D-吡喃半乳糖基)-(1 → 4)-(6-氧-乙酰基-苯-2-烯-α-D-赤型半乳糖基) 金雀异黄素，被命名为 G21，在分裂纺锤体中诱导畸变。在目前的研究中，我们探究了G21与遗传毒性活性相关的性质。用逆反应法和免疫带损耗法测试拓扑异构酶 IIα 活性的抑制，用免疫荧光法检测共价 DNA 拓扑异构酶 IIα 配合物和组蛋白 ɣH2AX。用微核化检测评估受试物的遗传毒性作用。用荧光标记法测试微核中着丝粒的存在和中心体的增殖。用分光光度法测定了微管蛋白聚合的抑制作用。我们发现这两种测试的药物都能够抑制拓扑异构酶II的活性；然而，G21，与金雀异黄素相比，在这种浓度对这种酶的阻滞远远超过具有细胞毒性的 IC(50)。我们还发现，这两种化合物均会造成DU 145 前列腺癌细胞微核化，但与金雀异黄素相比，G21 表现出细胞遗传毒性活性，由药物治疗细胞的微核中存在着丝粒来证明。G21的细胞遗传毒性产生自抑制微管蛋白聚合和中心体中断，在金雀异黄素存在时没有观察到。这项研究支持和扩展了我们以前的观测结果，金雀异黄素及其新型糖苷衍生物 G21的细胞毒性作用的机制明显不同。