解剖学报 ›› 2022, Vol. 53 ›› Issue (4): 507-514.doi: 10.16098/j.issn.0529-1356.2022.04.014

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Epsin3在结直肠癌中的差异表达及生物信息学分析

向丽莼1 姜钟翔2 姜小叶1 陈雪诺1 姜政1*
  

  1. 1. 重庆医科大学附属第一医院消化内科,重庆 400016;  2. 重庆市人民医院呼吸内科,重庆 400013
  • 收稿日期:2020-12-10 修回日期:2021-03-02 出版日期:2022-08-06 发布日期:2022-09-11
  • 通讯作者: 姜政 E-mail:zhengjiang1753@163.com

Differential expression and bioinformatics analysis of epsin3 in colorectal cancer

XIANG  Li-chun1  JIANG  Zhong-xiang2 JIANG  Xiao-ye1  CHEN  Xue-nuo1  JIANG  Zheng1*   

  1. Objective  To investigate the expression and significance of the adaptor protein epsin 3(EPN3) in colorectal cancer in order to provide reference for further study of EPN3.    Methods  GEPIA and GEDS were used to analyze the expression of EPN3 in colorectal cancer tissues and cells. SMART and cBioPortal databases were used to analyze the relationship between EPN3 gene methylation and copy number variation and its expression level. Metascape was used to complete analysis of gene ontology functional annotation and related pathways of EPN3 related genes and BioPlex was applied to construct a protein network in HCT116 cell. Thirteen pairs of colorectal cancer adjacent tissues and cancer tissue specimens were collected, and EPN3 mRNA expression were detected by Realtime PCR. The effect of abilities of cell proliferation, clone formation and migration via silencing EPN3 in HCT116 and HT29 were observed.    Results  GEPIA, GEDS, SMART and cBioPortal analyses showed that EPN3 was highly expressed in colorectal tumor tissues (P<0.01), and was related to methylation and copy number variation. The enrichment result  of EPN3 related genes showed that it was mainly related to cell adhesion. And a protein interaction network constructed by CCDC130, TNFAIP1, PHGDH, EPN2, etc. was related to protein ubiquitination. Real-time PCR result  showed that EPN3 was highly expressed in tumor tissues (P<0.05). Silencing EPN3 inhibited the proliferation, clony formation and migration abilities of HCT116 and HT29 cells.     Conclusion  EPN3 is highly expressed in colorectal cancer tissues and is related to cell adhesion and protein ubiquitination. Down-regulated EPN3 can inhibit abilities of proliferation, clony formation and migration of HCT116 and HT29 cells, which could provide a reference for further research on EPN3.
  • Received:2020-12-10 Revised:2021-03-02 Online:2022-08-06 Published:2022-09-11
  • Contact: JIANG Zheng E-mail:zhengjiang1753@163.com

摘要:

目的  探讨介导内吞作用的衔接蛋白epsin 3(EPN3)在结直肠癌中的表达及意义,为深入研究EPN3的调控模式提供实验依据。   方法  分别运用GEPIA和GEDS数据库分析EPN3在结直肠癌组织和细胞中的表达情况,并通过SMART和cBioPortal数据库分析EPN3基因甲基化和拷贝数变异与其表达水平的关系;利用Metascape数据库对EPN3相关的共表达基因集进行GO富集和通路分析;运用BioPlex蛋白互作数据库分析EPN3在HCT116细胞中的蛋白作用网络。为了对EPN3进一步验证,我们收集13对结直肠癌癌旁组织和癌组织标本,用Real-time PCR检测EPN3 mRNA表达;并通过敲减EPN3观察其对肿瘤细胞增殖、集落形成和迁移能力的影响。   结果  GEPIA、GEDS、SMART和cBioPortal等数据库分析显示,EPN3在结直肠肿瘤组织中高表达(P<0.01)。其表达水平与甲基化和拷贝数变异相关。EPN3相关基因的富集结果显示主要与细胞黏附相关。EPN3与UBB、CCDC130、TNFAIP1、PHGDH、EPN2等构成的蛋白相对作用网络与蛋白泛素化有关。Real-time PCR结果显示,EPN3在癌组织中高表达(P<0.05)。通过沉默EPN3可以抑制HCT116和HT29细胞的增殖、集落形成和迁移能力。   结论  EPN3在结直肠癌组织中高表达,且与细胞黏附和蛋白泛素化等生物学过程有关,敲低EPN3可抑制结直肠癌细胞系HCT116和HT29的增殖、集落形成和迁移等过程。

关键词: 结直肠癌, Epsin3, 生物信息学, 集落形成, 划痕实验, 实时定量聚合酶链反应,

Abstract: Objective To investigate the expression and significance of the adaptor protein EPN3 (Epsin 3) in colorectal cancer in order to provide reference for further study of EPN3. Methods We used tools of GEPIA and GEDS to analyze the expression of EPN3 in colorectal cancer tissues and cells. SMART and cBioPortal databases were used to analyze the relationship between EPN3 gene methylation and copy number variation and its expression level. Metascape was used to complete analysis of gene ontology (GO) functional annotation and related pathways of EPN3 related genes and BioPlex was applied to constructed a protein network in HCT116 cell. We collected 13 pairs of colorectal cancer adjacent tissue and cancer tissue specimens, and detected EPN3 mRNA expression by using RT-qPCR. We observed the effect of abilities of cell proliferation, clone formation and migration via silencing EPN3 in HCT116 and HT29. Results GEPIA, GEDS, SMART and cBioPortal analyses showed that EPN3 was highly expressed in colorectal tumor tissues (p<0.01), and was related to methylation and copy number variation. The enrichment results of EPN3 related genes showed that it was mainly related to cell adhesion. And a protein interaction network constructed by CCDC130, TNFAIP1, PHGDH, EPN2, etc. was related to protein ubiquitination. RT-qPCR result showed that EPN3 was highly expressed in tumor tissues (p<0.05). Silencing EPN3 inhibited the proliferation, clone formation and migration abilities of HCT116 and HT29 cells. Conclusion EPN3 is highly expressed in colorectal cancer tissues and is related to cell adhesion and protein ubiquitination. Down-regulated EPN3 can inhibit abilities of proliferation, clone formation and migration of HCT116 and HT29 cells, and this could provide a reference for further research on EPN3.

Key words: Colorectal cancer, Epsin3, Bioinformatics, Colony formation, Scratch test, Real-time PCR, Human

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