Yeast screening system reveals the inhibitory mechanism of cancer cell proliferation by benzyl isothiocyanate through down-regulation of Mis12
Yeast screening system reveals the inhibitory mechanism of cancer cell proliferation by benzyl isothiocyanate through down-regulation of Mis12
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DOI：
10.1038/s41598-019-45248-2
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发表时间：
2019-06-20
影响因子：
4.6
通讯作者：
Nakamura, Yoshimasa
中科院分区：
文献类型：
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作者：
Abe-Kanoh, Naomi;Kunisue, Narumi;Nakamura, Yoshimasa
异硫氰酸苄酯（BITC）是一种天然存在的异硫氰酸酯，来源于十字花科蔬菜。据报道，BITC可抑制多种癌细胞的增殖，这被认为对抑制肿瘤发生是重要的。然而，详细的作用机制仍不清楚。在这项研究中，我们采用了芽殖酵母酿酒酵母作为模式生物进行筛选。利用S.啤酒。动粒蛋白Mtw 1的过表达抵消了BITC在酵母中的抗增殖作用。BITC对人结肠癌HCT-116细胞增殖的抑制作用被Mtw 1的人直系同源物Mis 12的过表达所抑制，并被Mis 12的敲低所增强。我们还发现，BITC增加磷酸化和泛素化的Mis 12水平，从而减少Mis 12，表明BITC通过泛素-蛋白酶体系统降解Mis 12。细胞周期分析表明，Mis 12表达水平的改变影响了细胞周期分布和对BITC诱导凋亡的敏感性。这些结果提供的证据表明，BITC抑制细胞增殖，通过转录后调控的动粒蛋白Mis 12。
Benzyl isothiocyanate (BITC) is a naturally-occurring isothiocyanate derived from cruciferous vegetables. BITC has been reported to inhibit the proliferation of various cancer cells, which is believed to be important for the inhibition of tumorigenesis. However, the detailed mechanisms of action remain unclear. In this study, we employed a budding yeast Saccharomyces cerevisiae as a model organism for screening. Twelve genes including MTW1 were identified as the overexpression suppressors for the antiproliferative effect of BITC using the genome-wide multi-copy plasmid collection for S. cerevisiae. Overexpression of the kinetochore protein Mtw1 counteracts the antiproliferative effect of BITC in yeast. The inhibitory effect of BITC on the proliferation of human colon cancer HCT-116 cells was consistently suppressed by the overexpression of Mis12, a human orthologue of Mtw1, and enhanced by the knockdown of Mis12. We also found that BITC increased the phosphorylated and ubiquitinated Mis12 level with consequent reduction of Mis12, suggesting that BITC degrades Mis12 through an ubiquitin-proteasome system. Furthermore, cell cycle analysis showed that the change in the Mis12 level affected the cell cycle distribution and the sensitivity to the BITC-induced apoptosis. These results provide evidence that BITC suppresses cell proliferation through the post-transcriptional regulation of the kinetochore protein Mis12.