Xu Jian, Zhang Wei
Department of Hepatobiliary Surgery 1, Institute of Hepatobiliary-Pancreatic-Intestinal Diseases, Affiliated Hospital of North Sichuan Medical College, No. 1 Maoyuan nan Road, Shunqing District, Nanchang, 637000, China.
Department of Nuclear Medicine, Affiliated Hospital of North Sichuan Medical College, Nanchang, 637000, China.
Cancer Cell Int. 2021 Oct 9;21(1):521. doi: 10.1186/s12935-021-02222-1.
As a member of the ERM (ezrin-radixin-moesin) protein family, EZR has been recognized as a regulator of adhesion signal pathways by researchers. Moreover, EZR was thought to play irreplaceable roles in invasion and metastasis of versatile cancers. In this study, we managed to undermine the effect of EZR on proliferation and metastasis in pancreatic cancer (PC).
To analyze the impact of EZR expression on overall survival and free diseases survival of PC patients, we screened abnormally expressed EZR in PC using the Gene Expression Omnibus database (GEO database) and The Cancer Genome Atlas (TCGA) database. Following, Gene Ontology (GO)-based functional analysis and Gene set enrichment analysis (GSEA) was performed to predicate the possible biological processes in which EZR were involved. The clinicopathological characteristics and prognosis of PC patients were analyzed according to clinical data. Further, immunohistochemistry, western blotting and real time PCR analysis were conducted to analyze the expression level of EZR in PC and paired paracancerous tissues. The effect of EZR on proliferation of PC cell lines were detected by Cell Counting Kit-8 assay, and meanwhile, Transwell assay was performed to detect the effect of EZR on invasion and migration of PC cell.
EZR exhibited higher expression level in pancreatic cancer tissues and cell than paracancerous tissues and cell, and its expression level was positively correlated with poor overall survival and diseases-free survival in PC patients. CCK8 assay indicated that EZR facilitated the proliferation of PC cells, meanwhile, Transwell assay showed that EZR promoted the migration and invasion of PC cells. The GO analysis predicated that EZR was involved in biological processes including cell adhesion, ameboidal-type cell migration, cell junction assembly. Through GSEA analysis, pancreatic cancer pathway, and the adhesion junction pathway were screened as the mostly enriched pathways in EZR-regulated pathological process. The inhibition of EZR suppressed proliferation and migration of PC cells. Western blot experiment revealed a positive correlation between EZR and FAK, the proliferation invasion and migration ability of PC cells were significantly decreased after knockdown of EZR.
Our finding revealed EZR accelerated the progression of PC via FAK/AKT signaling pathway.
作为ERM（埃兹蛋白-根蛋白-膜突蛋白）蛋白家族的一员，EZR已被研究人员确认为黏附信号通路的调节因子。此外，EZR被认为在多种癌症的侵袭和转移中发挥着不可替代的作用。在本研究中，我们试图削弱EZR对胰腺癌（PC）增殖和转移的影响。
为了分析EZR表达对PC患者总生存期和无病生存期的影响，我们使用基因表达综合数据库（GEO数据库）和癌症基因组图谱（TCGA）数据库筛选PC中异常表达的EZR。随后，进行基于基因本体论（GO）的功能分析和基因集富集分析（GSEA），以预测EZR可能参与的生物学过程。根据临床数据分析PC患者的临床病理特征和预后。此外，进行免疫组织化学、蛋白质免疫印迹和实时PCR分析，以分析PC及配对癌旁组织中EZR的表达水平。通过细胞计数试剂盒-8法检测EZR对PC细胞系增殖的影响，同时进行Transwell实验检测EZR对PC细胞侵袭和迁移的影响。
EZR在胰腺癌组织和细胞中的表达水平高于癌旁组织和细胞，其表达水平与PC患者较差的总生存期和无病生存期呈正相关。CCK8实验表明EZR促进PC细胞增殖，同时，Transwell实验表明EZR促进PC细胞迁移和侵袭。GO分析预测EZR参与包括细胞黏附、阿米巴样细胞迁移、细胞连接组装等生物学过程。通过GSEA分析，胰腺癌通路和黏附连接通路被筛选为EZR调节的病理过程中最富集的通路。抑制EZR可抑制PC细胞的增殖和迁移。蛋白质免疫印迹实验显示EZR与FAK呈正相关，敲低EZR后PC细胞的增殖侵袭和迁移能力显著降低。
我们的研究结果表明EZR通过FAK/AKT信号通路加速了PC的进展。
