Nek2 augments sorafenib resistance by regulating the ubiquitination and localization of β-catenin in hepatocellular carcinoma
Nek2 augments sorafenib resistance by regulating the ubiquitination and localization of β-catenin in hepatocellular carcinoma
复制标题
Nek2 通过调节肝细胞癌中 β-连环蛋白的泛素化和定位来增强索拉非尼耐药性
DOI：
10.1186/s13046-019-1311-z
复制
发表时间：
2019-07-18
影响因子：
11.3
通讯作者：
Wu, Dehua
中科院分区：
文献类型：
--
作者：
Deng, Ling;Sun, Jingyuan;Wu, Dehua
背景：索拉非尼是晚期肝细胞癌（HCC）的一线治疗药物。一些研究表明，β-连环蛋白的上调在肝癌索拉非尼耐药中起作用;然而，与这种现象相关的机制仍然habitu.Methods：Western印迹，流式细胞术，和IC 50值的评价被用来确认β-连环蛋白在肝癌索拉非尼耐药中的作用。然后进行免疫沉淀和蛋白质印迹以鉴定β-连环蛋白和Nek 2之间的调节相互作用。此外，使用蛋白质印迹、流式细胞术和体内异种移植模型来评估Nek 2在HCC索拉非尼抗性中的功能，而进行拯救实验以证实Nek 2通过β-连环蛋白诱导索拉非尼抗性。最后，采用免疫印迹法和免疫组化法检测Nek 2在肝癌和癌旁组织中的表达水平。结果：我们发现β-catenin可以抑制索拉非尼诱导的肝癌细胞凋亡和细胞生长抑制。通过筛选β-连环蛋白相互作用蛋白，我们发现Nek 2可以在索拉非尼处理的HCC细胞系中结合β-连环蛋白。我们的研究结果还表明，Nek 2稳定β-连环蛋白，并促进其易位到细胞核，从而激活下游靶基因的转录。我们进一步证实了Nek 2可以诱导肝癌细胞系对索拉非尼产生耐药性，而β-连环蛋白是参与这一过程的关键因素。此外，异种移植肿瘤模型显示Nek 2敲低可以改善索拉非尼的抗肿瘤作用，而肿瘤蛋白的分析显示Nek 2调节体内β-连环蛋白蛋白水平及其核转位。此外，Nek 2被发现在肝癌组织中上调，尤其是在晚期diseases.Conclusions：我们的研究证明，Nek 2诱导肝癌索拉非尼耐药通过β-连环蛋白，并提出了一种新的治疗策略，以提高索拉非尼在肝癌的抗肿瘤作用。
Background: Sorafenib is the first-line treatment for advanced-stage hepatocellular carcinoma (HCC). Several studies have shown that the up-regulation of beta-catenin plays a role in sorafenib resistance in HCC; however, the mechanism associated with this phenomenon remains elusive.Methods: Western blotting, flow cytometry, and an evaluation of IC50 values were used to confirm the role of beta-catenin in HCC sorafenib resistance. Immunoprecipitation and western blotting were then performed to identify regulatory interactions between beta-catenin and Nek2. Further, western blotting, flow cytometry, and an in vivo xenograft model were used to evaluate the function of Nek2 in HCC sorafenib resistance, whereas rescue experiments were performed to confirm that Nek2 induces sorafenib resistance via beta-catenin. Finally, western blotting and immunohistochemistry were used to evaluate the expression level of Nek2 in paired HCC and non-tumor tissues.Results: We showed that beta-catenin could suppress sorafenib-induced apoptosis and cell growth inhibition in HCC cell lines. By screening beta-catenin-interacting proteins, we found that Nek2 could bind beta-catenin in sorafenib-treated HCC cell lines. Our results also showed that Nek2 stabilizes beta-catenin and promotes its translocation to the nucleus, consequently activating the transcription of downstream target genes. We further confirmed that Nek2 could induce sorafenib resistance in HCC cell lines, and that beta-catenin was the key element involved in this process. Further, a xenograft tumor model showed that Nek2 knockdown could improve the anti-tumor effect of sorafenib, whereas an analysis of tumor proteins showed that Nek2 regulates beta-catenin protein levels and its nuclear translocation in vivo. In addition, Nek2 was found to be up-regulated in HCC tissue, and especially in advanced-stage disease.Conclusions: Our study proves that Nek2 induces HCC sorafenib resistance via beta-catenin and suggests a novel therapeutic strategy to improve the anti-tumor effects of sorafenib in HCC.