The direct interaction between Cullin 4B (CUL4B) and DNA damage-binding protein 1 (DDB1) is required for the assembly of Cullin4B-RING E3 ligase complex (CRL4B), which are involved in the tumorigenesis of osteosarcoma through ubiquitinating and degrading multiple tumor suppressors and cell cycle regulators. Thus, targeting CUL4B-DDB1 interaction to prevent the assembly of CRL4B may be a potent approach to inhibit osteosarcoma cell growth. In the present study, we identified six naturally-sourced small molecules that can specifically disrupt the CUL4B-DDB1 interaction using an in vitro high-throughput screening (HTS) system in yeast. We focused our investigation on revealing the molecular effects of TSC01682, the most active compound capable of inhibiting osteosarcoma cell growth. Biochemically, TSC01682 significantly repressed the CUL4B-DDB1 interaction in both yeast cells and osteosarcoma cells. Moreover, TSC01682 treatment in osteosarcoma cells also caused a decrease of other CRL4B components including CUL4-associated factor 11 (DCAF11) and DCAF13, but an increase of two CRL4B substrates including cyclin-dependent kinase inhibitor 1A (CDKN1A, also known as p21) and phosphatase and tensin homolog deleted on chromosome 10 (PTEN) through inhibiting their ubiquitination. Consistent with these molecular changes, TSC01682 treatment significantly inhibited cell proliferation, colony formation, invasion, and in vivo tumor growth. Collectively, our results suggest that TSC01682 is a potent compound capable of disrupting the CUL4B-DDB1 interaction, and it may be developed as a chemotherapeutic drug for osteosarcoma treatment.

译文

:Cullin4B-RING E3连接酶复合物(CRL4B)的组装需要Cullin 4B(CUL4B)和DNA损伤结合蛋白1(DDB1)之间的直接相互作用,而Cullin4B-RING E3连接酶复合物(CRL4B)通过泛素化和降解多个肿瘤参与骨肉瘤的肿瘤发生抑制剂和细胞周期调节剂。因此,靶向CUL4B-DDB1相互作用以防止CRL4B的组装可能是抑制骨肉瘤细胞生长的有效方法。在本研究中,我们鉴定了六个天然来源的小分子,它们可以使用酵母中的体外高通量筛选(HTS)系统特异性破坏CUL4B-DDB1的相互作用。我们的研究集中在揭示TSC01682的分子作用,TSC01682是能够抑制骨肉瘤细胞生长的最具活性的化合物。生化方面,TSC01682显着抑制酵母细胞和骨肉瘤细胞中的CUL4B-DDB1相互作用。此外,在骨肉瘤细胞中进行TSC01682处理还导致其他CRL4B组分(包括CUL4相关因子11(DCAF11)和DCAF13)减少,但增加了两种CRL4B底物,包括细胞周期蛋白依赖性激酶抑制剂1A(CDKN1A,也称为p21)和磷酸酶和张力蛋白同源物通过抑制它们的泛素化作用而在第10号染色体(PTEN)上缺失。与这些分子变化一致,TSC01682处理可显着抑制细胞增殖,集落形成,侵袭和体内肿瘤生长。总的来说,我们的结果表明,TSC01682是一种能够破坏CUL4B-DDB1相互作用的有效化合物,它可能被开发为用于骨肉瘤治疗的化疗药物。

+1
+2
100研值 100研值 ¥99课程
检索文献一次
下载文献一次

去下载>

成功解锁2个技能,为你点赞

《SCI写作十大必备语法》
解决你的SCI语法难题!

技能熟练度+1

视频课《玩转文献检索》
让你成为检索达人!

恭喜完成新手挑战

手机微信扫一扫,添加好友领取

免费领《Endnote文献管理工具+教程》

微信扫码, 免费领取

手机登录

获取验证码
登录