NCSTN promotes hepatocellular carcinoma cell growth and metastasis via β-catenin activation in a Notch1/AKT dependent manner
NCSTN promotes hepatocellular carcinoma cell growth and metastasis via β-catenin activation in a Notch1/AKT dependent manner
复制标题
NCSTN 以 Notch1/AKT 依赖性方式通过 β-catenin 激活促进肝细胞癌细胞生长和转移
DOI：
10.1186/s13046-020-01638-3
复制
发表时间：
2020-07-06
影响因子：
11.3
通讯作者：
Wu, Hong
中科院分区：
文献类型：
--
作者：
Li, Hui;Lan, Tian;Wu, Hong
背景：肝细胞癌是全球癌症相关死亡的第三大原因。HCC进展迅速，复发率高，预后较差。Nicastrin （NCSTN）是γ -Secretase的核心亚基，据报道在肿瘤进展中起重要作用。然而，目前尚无研究揭示其在HCC中的作用。方法采用免疫组织化学染色、Western blot和实时荧光定量PCR检测NCSTN的表达。细胞计数试剂盒-8，菌落形成和细胞周期测定法用于体外评估细胞生长。采用Transwell法和创面愈合法评估细胞迁移和侵袭能力。采用免疫荧光、亚细胞蛋白分离和共免疫沉淀法对β -catenin进行定位分析。异种移植肿瘤模型证实了NCSTN的体内功能。结果与正常肝组织相比，sncstn在HCC中显著过表达。升高的NCSTN表达水平与HCC患者的总生存率和无复发生存率显著相关。NCSTN表达增强可促进肝癌细胞在体内外的生长、迁移和侵袭。机制研究表明NCSTN通过上调Zeb1诱导上皮-间质转化（EMT）过程。随后，我们发现NCSTN促进了β -catenin的核易位，β -catenin是Zeb1的正转录调节因子。利用Notch和AKT抑制剂，我们发现NCSTN通过Notch1和AKT信号通路促进β -catenin的激活。NCSTN通过切割Notch1增加AKT和GSK-3 β的磷酸化，从而降低GSK-3 β / β -catenin复合物。GSK-3 β的失活抑制了β -catenin的降解，促进β -catenin的核易位启动Zeb1的转录，导致恶性表型。结论NCSTN通过β -catenin介导的Zeb1的上调，以Notch1/AKT依赖的方式促进HCC细胞的生长和转移，提示NCSTN可能是HCC的潜在预后标志物和治疗靶点。
BackgroundHepatocellular carcinoma is the third top cause of cancer-related mortalities worldwide. The prognosis of HCC patients remains poor due to rapid progression and high incidence of tumor recurrence. Nicastrin (NCSTN), a core subunit of gamma -Secretase, has been reported to play a vital role in tumor progression. However, no study till now has revealed its role in HCC.MethodsThe expression of NCSTN was evaluated by immunohistochemical staining, Western blot, and quantitative real-time PCR. Cell counting kit-8, colony formation and cell cycle assays were used for evaluating cell growth in vitro. Transwell and wound-healing assays were used for evaluating cell migration and invasion capacity. Immunofluorescence, subcellular protein fractionation and co-immunoprecipitation were used for location analysis of beta -catenin. The in vivo functions of NCSTN were illustrated by xenograft tumor models.ResultsNCSTN was dramatically overexpressed in HCC compared to normal liver tissues. Elevated NCSTN expression level was significantly correlated to worse overall and recurrence-free survival of HCC patients. Enhanced NCSTN expression promoted HCC cell growth, migration and invasion in vitro and in vivo. Mechanistic investigations showed that NCSTN induced epithelial-mesenchymal transition (EMT) process via upregulation of Zeb1. Subsequently, we revealed that NCSTN facilitated nuclear translocation of beta -catenin, a positive transcriptional regulator of Zeb1. Using Notch and AKT inhibitors, we revealed that NCSTN promoted beta -catenin activation through Notch1 and AKT signaling pathway. NCSTN increased AKT and GSK-3 beta phosphorylation by cleavage of Notch1, which decreased GSK-3 beta/beta -catenin complex. The inactivation of GSK-3 beta inhibited the beta -catenin degradation and promoted nuclear translocation of beta -catenin to initiate transcription of Zeb1, resulting in malignant phenotype.ConclusionsOur results demonstrated that NCSTN promoted HCC cell growth and metastasis via beta -catenin-mediated upregulation of Zeb1 in a Notch1/AKT dependent manner, suggesting that NCSTN might serve as a potential prognostic marker and therapeutic target for HCC.