NEK2 promotes the migration and proliferation of ESCC via stabilization of YAP1 by phosphorylation at Thr-143.
NEK2 promotes the migration and proliferation of ESCC via stabilization of YAP1 by phosphorylation at Thr-143.
复制标题
DOI：
10.1186/s12964-022-00898-0
复制
发表时间：
2022-06-15
期刊：
影响因子：
--
通讯作者：
中科院分区：
文献类型：
--
作者：
食管鳞状细胞癌（ESCC）是一种区域性、侵袭性、高发病率和高死亡率的肿瘤。NIMA相关激酶2（NEK 2）是一种有趣的癌基因，其改变导致患者受益的结果。本研究旨在探讨NEK 2在食管鳞癌中的作用及其机制。从TCGA和GEO下载RNA-seq数据并通过R软件分析。采用免疫组化（IHC）或蛋白质印迹（WB）检测蛋白质水平，qRT-PCR检测mRNA表达。采用Transwell迁移实验和集落形成实验分别检测细胞的体外增殖和迁移作用。使用皮下异种移植肿瘤模型探索体内作用，其中采用免疫荧光（IF）和IHC研究表达和定位。免疫沉淀法检测蛋白质间的相互作用。在放线菌酮存在下，通过WB测定蛋白质的稳定性。GEO、TCGA和组织芯片检测结果显示，食管鳞癌组织中NEK 2的表达水平高于正常食管上皮，且与食管癌的严重程度相关。NEK 2基因敲低可抑制食管鳞癌细胞的增殖和迁移，并下调YAP 1和EMT标志物如N-cadherin和Vimentin的表达。相反，NEK 2过表达增强了ESCC的迁移，并升高了YAP 1、N-cadherin和Vimentin的水平。此外，NEK 2中YAP 1的过表达敲低了ESCC，部分挽救了相应的迁移减少。NEK 2的敲低在体内发挥抗肿瘤作用，并伴随着YAP 1的低水平和核改组。在机制上，NEK 2与YAP 1相互作用，通过阻止泛素化增加内源性和外源性YAP 1的稳定性。此外，计算机预测的YAP 1的磷酸化位点，Thr-143，减少HA-YAP 1的泛素化，增强其稳定性，从而影响体外迁移。NEK 2是一种在ESCC中高表达的预后癌基因，在体外和体内促进ESCC的进展。从机制上讲，NEK 2介导的YAP 1在Thr-143的磷酸化保护其免受蛋白酶体降解，并可能成为ESCC中有希望的治疗靶点。视频摘要在线版本包含补充材料，可通过10. 1186/s12964-022-00898-0获取。
Esophageal Squamous Cell Carcinoma (ESCC) was characterized as a regional-prevalent and aggressive tumor with high morbidity and mortality. NIMA-related kinase 2 (NEK2) is an interesting oncogene, the alteration of which leads to patients-beneficial outcomes. We aimed to explore the role of NEK2 in ESCC and excavate its mechanism. RNA-seq data were downloaded from TCGA and GEO and analyzed by R software. The protein levels were detected by immunohistochemistry (IHC) or western blot (WB), and mRNA expression was detected by qRT-PCR. The in vitro role of proliferation and migration was detected by Transwell migration assay and by colony formation assay, respectively. The in vivo roles were explored using a subcutaneous xenograft tumor model, where immunofluorescence (IF) and IHC were employed to investigate expression and localization. The interaction between proteins was detected by immunoprecipitation. The stability of proteins was measured by WB in the presence of cycloheximide. A higher level of NEK2 was found in ESCC than normal esophageal epithelia in GEO, TCGA, and tissue microarray, which was associated with worse prognoses. The NEK2 knockdown impaired the proliferation and migration of ESCC, which also downregulated YAP1 and EMT markers like N-cadherin and Vimentin in vitro. On the contrary, NEK2 overexpression enhanced the migration of ESCC and elevated the levels of YAP1, N-cadherin, and Vimentin. Additionally, the overexpression of YAP1 in NEK2 knocked down ESCCs partly rescued the corresponding decrease in migration. The knockdown of NEK2 played an anti-tumor role in vivo and was accompanied by a lower level and nucleus shuffling of YAP1. In mechanism, NEK2 interacted with YAP1 and increased the stability of both endogenous and exogenous YAP1 by preventing ubiquitination. Moreover, the computer-predicted phosphorylation site of YAP1, Thr-143, reduced the ubiquitination of HA-YAP1, strengthened its stability, and thus influenced the migration in vitro. NEK2 is a prognostic oncogene highly expressed in ESCC and promotes the progression of ESCC in vitro and in vivo. Mechanistically, NEK2-mediated phosphorylation of YAP1 at Thr-143 protects it from proteasome degradation and might serve as a promising therapeutic target in ESCC. Video Abstract The online version contains supplementary material available at 10.1186/s12964-022-00898-0.
登录
查看更多内容
不平衡的 YAP-SOX9 回路驱动食管鳞状细胞癌的干细胞性和恶性进展
影响因子：
8
作者：
Wang L;Zhang Z;Yu X;Huang X;Liu Z;Chai Y;Yang L;Wang Q;Li M;Zhao J;Hou J;Li F
通讯作者：
Li F
NEK2磷酸化并稳定β-catenin在PLK1下游的有丝分裂中心体上。
影响因子：
3.3
作者：
Mbom BC;Siemers KA;Ostrowski MA;Nelson WJ;Barth AI
通讯作者：
Barth AI
影响因子：
14.9
作者：
Naro C;Barbagallo F;Chieffi P;Bourgeois CF;Paronetto MP;Sette C
通讯作者：
Sette C
影响因子：
5.6
作者：
Wang, Chongchong;Cheng, Li;Sun, Guoping
通讯作者：
Sun, Guoping
DOI：
10.1016/s0140-6736(17)33326-3
发表时间：
2018-03-17
期刊：
Lancet (London, England)
影响因子：
--
作者：
Allemani C;Matsuda T;Di Carlo V;Harewood R;Matz M;Nikšić M;Bonaventure A;Valkov M;Johnson CJ;Estève J;Ogunbiyi OJ;Azevedo E Silva G;Chen WQ;Eser S;Engholm G;Stiller CA;Monnereau A;Woods RR;Visser O;Lim GH;Aitken J;Weir HK;Coleman MP;CONCORD Working Group
通讯作者：
CONCORD Working Group