Extract of bulbus of Fritillaria cirrhosa induces spindle multipolarity in human-derived colonic epithelial NCM460 cells through promoting centrosome fragmentation
Extract of bulbus of Fritillaria cirrhosa induces spindle multipolarity in human-derived colonic epithelial NCM460 cells through promoting centrosome fragmentation
复制标题
川贝母提取物通过促进中心体破碎诱导人源性结肠上皮 NCM460 细胞纺锤体多极性
DOI：
10.1093/mutage/geab002
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发表时间：
2021
期刊：
影响因子：
2.7
通讯作者：
Wang Xu
中科院分区：
文献类型：
--
作者：
Guo Xihan;Wang Chunlei;Tian Weimeng;Dai Xueqin;Ni Juan;Wu Xiayu;Wang Xu
浙贝母Don（BFC）是一种在中国和许多其他国家使用的优秀止咳祛痰中药，具有潜在的遗传毒性，但了解较少。以前，我们已经报道了BFC的水提取物在NCM 460细胞中损害纺锤体组装检查点和胞质分裂。在这里，我们发现，一个显着的观察，在BFC处理的NCM 460细胞是多极有丝分裂，一个性状的经典妥协的保真度染色体分离。更详细的研究表明，BFC诱导的纺锤体多极化的中期和后期的剂量和时间依赖性的方式，表明BFC诱导的多极纺锤体构象不是短暂的。多极中期的频率与多极间末期的频率相关性很好，表明BFC诱导的多极中期经常持续到后期。出乎意料的是，BFC阻断了双核细胞的增殖，表明纺锤体多极化不是BFC诱导的胞质分裂失败的下游。BFC暴露于早期有丝分裂细胞，而不是S/G2细胞，极大地促进了纺锤体多极化，表明BFC可能破坏中心体的完整性，而不是诱导中心体过度复制。免疫荧光结果显示，BFC短期处理后，中心体断裂严重，有丝分裂早期细胞的中心体断裂程度大于S/G2细胞。与此同时，BFC对一些参与调节中心体完整性的基因（如p53、Rb centrin-2、Plk-4、Plk-1和Aurora-A）的调控也发生了显著的下调。总之，我们的研究结果表明，BFC导致多极纺锤体主要是通过诱导中心体碎片。将这些结果与我们以前的观察结果相结合，我们建议在实际使用BFC时应考虑风险/受益比。
Bulbus ofFritillaria cirrhosaD. Don (BFC), an outstanding antitussive and expectorant herbal drug used in China and many other countries, has potential but less understood genotoxicity. Previously, we have reported that aqueous extract of BFC compromised the spindle assembly checkpoint and cytokinesis in NCM460 cells. Here, we found that one remarkable observation in BFC-treated NCM460 cells was multipolar mitosis, a trait classically compromises the fidelity of chromosome segregation. More detailed investigation revealed that BFC-induced spindle multipolarity in metaphases and ana-telophases in a dose- and time-dependent manner, suggesting BFC-induced multipolar spindle conformation was not transient. The frequency of multipolar metaphase correlated well to that of multipolar ana-telophases, indicating that BFC-induced multipolar metaphases often persisted through anaphase. Unexpectedly, BFC blocked the proliferation of binucleated cells, suggesting spindle multipolarity was not downstream of BFC-induced cytokinesis failure. Exposure of BFC to early mitotic cells, rather than S/G2 cells, contributed greatly to spindle multipolarity, indicating BFC might disrupt centrosome integrity rather than induce centrosome overduplication. The immunofluorescence results showed that the centrosomes were severely fragmented by a short-term treatment of BFC and the extent of centrosome fragmentation in early mitotic cells was larger than this in S/G2 cells. Consistently, several genes (e.g.p53,Rb centrin-2,Plk-4,Plk-1andAurora-A) involved in regulating centrosome integrity were significantly deregulated by BFC. Together, our results suggest that BFC causes multipolar spindles primarily by inducing centrosome fragmentation. Coupling these results to our previous observations, we recommend the risk/benefit ratio should be considered in the practical use of BFC.