NEK2 promotes the migration and proliferation of ESCC via stabilization of YAP1 by phosphorylation at Thr-143

Abstract

Background 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. Methods 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. Results 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. Conclusions 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.

Differential Expression of NEK Kinase Family Members in Esophageal Adenocarcinoma and Barrett’s Esophagus

“…NEK2 overexpression has been found in various human malignancies [73][74][75][76][77][78][79][80], including gastrointestinal tumors [81,82]. Aberrant NEK2 expression in cancers has been associated with tumor cell proliferation [83], migration [82], invasion [74,77], drug/radio resistance [84,85], immune response [80,86], and poor prognosis [26,75]. In our bioinformatics analyses, NEK2 is the most frequently upregulated NEK member in EAC (significantly detected in seven of the eight databases).…”

“…In addition, NEK2 may also play a role in microtubule stabilization [70], B cell development and immunological response [71], and splicing [72]. NEK2 overexpression has been found in various human malignancies [73][74][75][76][77][78][79][80], including gastrointestinal tumors [81,82]. Aberrant NEK2 expression in cancers has been associated with tumor cell proliferation [83], migration [82], invasion [74,77], drug/radio resistance [84,85], immune response [80,86], and poor prognosis [26,75].…”

NEK2 promotes TP53 ubiquitination to enhance the proliferation and migration of TP53 wild-type glioblastoma cells

“…The NIMA-related kinase (NEK) 2, a serine/threonine kinase belonging to the NEK family, exhibits specific localization at the centrosome. It is widely acknowledged as a multifunctional protein involved in regulating various aspects of the cell cycle [6][7][8][9][10]. NEK2 as multiple research studies have consistently observed its increased expression in different types of cancers and its correlation with poor overall survival results [11][12][13][14].…”

“…The NIMA-related kinase (NEK) 2, a serine/threonine kinase belonging to the NEK family, exhibits specific localization at the centrosome [6]. NEK2 is widely acknowledged as a versatile protein involved in the regulation of various aspects of cell cycle, including replication and segregation of centrosomes, stabilization of microtubules, attachment of kinetochores, and activation of the spindle assembly checkpoint [7][8][9][10]. In the realm of scientific academia, there has been considerable focus on the significant involvement of NEK2 in the development of tumors in recent years.…”

Exploring the hub genes and mechanisms of Daphne altaica treating esophageal squamous cell carcinoma based on network pharmacology and bioinformatics analysis

“…2021) Never in mitosis-related kinase 2 (NEK2) is a serine/threonine kinase that facilitates the division of centrosomes and guarantees the accurate segregation of chromosomes in the G2/M phase of the cell cycle(Xia et al 2015). The prognostic signi cance of increased NEK2 expression and its role in ESCC in proliferation, migration, and EMT was demonstrated in a study by Su W et al They concluded that NEK2 interacts with YAP1 and phosphorylates it at Thr-143, thereby attenuating its ubiquitylation in vitro and promoting its stability, thereby mediating the progression of ESCC(Su et al 2022). Previous studies have indicated that NEK2 inhibitors e ciently inhibit cancer cell proliferation by causing arrest of the cellular cycle and apoptotic cell death, and markedly inhibition of tumor growth in-vivo(Xi et al 2017;Fang et al 2016).…”