Laboratory of Cell Biophysics, Division of Molecular Biology, Ruđer Bošković Institute, Zagreb 10000, Croatia.
Proc Natl Acad Sci U S A. 2024 Nov 19;121(47):e2405015121. doi: 10.1073/pnas.2405015121. Epub 2024 Nov 14.
Cancer cells often display errors in chromosome segregation, some of which result from improper chromosome alignment at the spindle midplane. Chromosome alignment is facilitated by different rates of microtubule poleward flux between sister kinetochore fibers. However, the role of the poleward flux in supporting mitotic fidelity remains unknown. Here, we introduce the hypothesis that the finely tuned poleward flux safeguards against lagging chromosomes and micronuclei at mitotic exit by promoting chromosome alignment in metaphase. We used human untransformed RPE-1 cells depleted of KIF18A/kinesin-8 as a system with reduced mitotic fidelity, which we rescued by three mechanistically independent treatments, comprising low-dose taxol or codepletion of the spindle proteins HAUS8 or NuMA. The rescue of mitotic errors was due to shortening of the excessively long overlaps of antiparallel microtubules, serving as a platform for motor proteins that drive the flux, which in turn slowed down the overly fast flux and improved chromosome alignment. In contrast to the prevailing view, the rescue was not accompanied by reduction of overall microtubule growth rates. Instead, speckle microscopy revealed that the improved chromosome alignment in the rescue treatments was associated with slower growth and flux of kinetochore microtubules. In a similar manner, a low-dose taxol treatment rescued mitotic errors in a high-grade serous ovarian carcinoma cell line OVKATE. Collectively, our results highlight the potential of targeting microtubule poleward flux to modify chromosome instability and provide insight into the mechanism through which low doses of taxol rescue certain mitotic errors in cancer cells.
癌细胞经常在染色体分离过程中出现错误,其中一些错误是由于纺锤体中部的姐妹动粒纤维之间微管极向流速的不同而导致的。然而,极向流速在支持有丝分裂保真度方面的作用仍然未知。在这里,我们提出了一个假设,即精细调节的极向流速通过促进中期染色体的对齐来防止滞后染色体和有丝分裂后期的微核。我们使用人类未转化的 RPE-1 细胞,通过敲除 KIF18A/驱动蛋白-8 作为一个具有降低有丝分裂保真度的系统,我们通过三种机制上独立的处理来挽救这个系统,包括低剂量紫杉醇或纺锤体蛋白 HAUS8 或 NuMA 的共缺失。有丝分裂错误的挽救是由于过长的平行微管重叠的缩短,这为驱动流速的马达蛋白提供了一个平台,从而减缓了过快的流速并改善了染色体的对齐。与流行的观点相反,挽救并没有伴随着整体微管生长速度的降低。相反,斑点显微镜显示,挽救处理中改善的染色体对齐与动粒微管的生长和流速较慢有关。以类似的方式,低剂量紫杉醇处理挽救了高级别浆液性卵巢癌细胞系 OVKATE 中的有丝分裂错误。总的来说,我们的结果强调了靶向微管极向流速以改变染色体不稳定性的潜力,并深入了解了低剂量紫杉醇挽救癌细胞中某些有丝分裂错误的机制。