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
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NCSTN 通过 β-catenin 以 Notch1/AKT 依赖性方式激活促进肝细胞癌细胞生长和转移
DOI：
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发表时间：
2020
影响因子：
11.3
通讯作者：
Wu Hong
中科院分区：
文献类型：
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作者：
Li Hui;Lan Tian;Xu Lin;Liu Hailing;Wang Jinjv;Li Jiaxin;Chen Xiangzheng;Huang Jiwei;Yuan Kefei;Zeng Yong;Wu Hong
背景：肝细胞癌是全球第三大癌症相关死亡原因。肝癌患者的预后仍然很差，因为肿瘤进展快，复发率高。尼卡斯特林已报道γ-分泌酶的核心亚基（NCBI）在肿瘤进展中起重要作用。然而，no.study到目前为止已经揭示了它在HCC中的作用。方法：采用免疫组化、Western blot和实时荧光定量PCR检测NCBI的表达。采用细胞计数试剂盒-8、集落形成和细胞周期测定法评价细胞体外生长情况。Transwell和伤口愈合测定用于评估细胞迁移和侵袭能力。采用免疫荧光、亚细胞蛋白分级和免疫共沉淀法对β-catenin进行定位分析。通过异种移植肿瘤模型说明了NCBI的体内功能。结果：与正常肝组织相比，HCC中NCBI显著过表达。NCBI表达水平升高与HCC患者总体生存率和无复发生存率降低显著相关。增强的NCBI表达促进肝癌细胞的生长、迁移和侵袭。机制研究表明，NCBI通过上调.Zeb1诱导上皮-间质转化（EMT）过程。随后，我们发现NCBI促进了β-catenin的核转位，β-catenin是Zeb 1的正性转录调节因子。使用Notch和AKT抑制剂，我们发现NCBI通过Notch 1和AKT信号通路促进β-catenin活化。NCBI通过切割Notch 1增加AKT和GSK-3β的磷酸化，从而降低GSK-3β/β-catenin复合物。GSK-3β的失活抑制了β-catenin的降解，促进了β-catenin的核转位，启动了Zeb 1的转录，导致恶性表型。结论：我们的研究结果表明，NCBI通过β-catenin介导的Notch 1/AKT依赖性上调Zeb 1的表达促进HCC细胞的生长和转移，提示NCBI可能作为HCC的潜在预后标志物和治疗靶点。
Background: Hepatocellular 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 γ-Secretase, has been reported to play a vital role in tumor progression. However, no.study till now has revealed its role in HCC..Methods: The 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 β-catenin. The in vivo functions of NCSTN were illustrated by xenograft tumor models..Results: NCSTN 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 β-catenin, a positive transcriptional.regulator of Zeb1. Using Notch and AKT inhibitors, we revealed that NCSTN promoted β-catenin activation through.Notch1 and AKT signaling pathway. NCSTN increased AKT and GSK-3β phosphorylation by cleavage of Notch1,.which decreased GSK-3β/β-catenin complex. The inactivation of GSK-3β inhibited the β-catenin degradation and.promoted nuclear translocation of β-catenin to initiate transcription of Zeb1, resulting in malignant phenotype..Conclusions: Our results demonstrated that NCSTN promoted HCC cell growth and metastasis via β-cateninmediated.upregulation of Zeb1 in a Notch1/AKT dependent manner, suggesting that NCSTN might serve as a potential prognostic marker and therapeutic target for HCC.