食管胃结合部腺癌生物信息学分析及Involucrin基因突变位点筛选

doi:10.16352/j.issn.1001-6325.2022.02.028

基础医学与临床 ›› 2022, Vol. 42 ›› Issue (2): 235-242.doi: 10.16352/j.issn.1001-6325.2022.02.028

食管胃结合部腺癌生物信息学分析及Involucrin基因突变位点筛选

吕雪¹, 李雪薇¹, 杨婷¹, 郑锦秀², 祝子鹤¹, 杨涛^1,3*, 徐钧^4*

山西医科大学 1.生物化学与分子生物学教研室;
2.药理学教研室;
3.细胞生理学教育部重点实验室,山西太原 030001;
4.山西医科大学第一医院肝胆胰外科,山西太原 030001

收稿日期:2021-02-24 修回日期:2021-06-13 出版日期:2022-02-05 发布日期:2022-01-24
通讯作者: ^* yangtao056@126.com;junxuty@163.com
基金资助:
国家自然科学基金(82072737)

Bioinformatic analysis of adenocarcinoma at esophagogastric junction and mutation sites screening of Involucrin gene

LYU Xue¹, LI Xue-wei¹, YANG Ting¹, ZHENG Jin-xiu², ZHU Zi-he¹, YANG Tao^1,3*, XU Jun^4*

Department of Biochemistry and Molecular Biology;
2. Department of Pharmacology;
3. Key Laboratory of Cellular Physiology,Ministry of Education, Shanxi Medical University,Taiyuan 030001;
4. Department of Hepatopancreatobiliary Surgery,the First Hospital of Shanxi Medical University, Taiyuan 030001,China

Received:2021-02-24 Revised:2021-06-13 Online:2022-02-05 Published:2022-01-24
Contact: ^* yangtao056@126.com;junxuty@163.com

摘要/Abstract

摘要： 目的检测食管胃结合部腺癌(AEG)患者基因突变情况,分析内披蛋白(IVL)突变对肿瘤细胞生物学功能的影响。方法将22例山西地区食管胃结合部腺癌患者的癌组织和癌旁组织进行全外显子组测序,分析IVL的突变情况并筛选IVL突变位点;整理汇总TCGA数据库中食管癌IVL的突变情况,进行GO功能注释和KEGG富集分析。构建IVL过表达质粒,转染食管癌细胞系KYSE150,测定细胞的增殖和迁移能力。体外合成不同突变位点的IVL寡肽,通过体外活性测试分析寡肽与转谷氨酰胺酶1(TGase1)的结合能力。结果测序结果发现在食管胃结合部腺癌中IVL有5处突变;GO分析显示IVL突变与上皮细胞分化、细胞分化调控的活性等生物学过程有关;KEGG信号通路分析显示IVL突变与cAMP信号通路等途径有关。过表达IVL可抑制食管癌细胞的增殖和迁移能力(P＜0.05)。突变型IVL寡肽与TGase1的结合能力弱于野生型寡肽(P＜0.05)。结论 IVL重复基序第7位氨基酸的突变减弱IVL与TGase1的结合,影响多种与肿瘤相关的生物学过程和信号通路。

关键词: 食管胃结合部腺癌, 内披蛋白, 基因突变

Abstract: Objective To detect the gene mutations in patients with Adenocarcinoma at esophagogastric junction(AEG) and to analyze the effect of Involucrin (IVL) mutation on the biological function of tumor cells. Methods Whole exome sequencing was performed on cancerous and paired adjacent normal tissues from 22 AEG patients in Shanxi province. IVL was selected for further investigation of mutation sites. Esophagus cancer data from TCGA were divided into three groups based on IVL mutations. Differentially-expressed genes were screened and proceeded via Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Cell proliferation and migration assays were performed to evaluate the effect of over-expression of IVL on KYSE 150 cells. Mutant IVL oligopeptides were synthesized and used to evaluate the binding activity with TGase1. Results Five IVL mutation sites were identified in 22 AEG. GO annotation indicated that these mutations were related to the epithelial differentiation and activity of cell differentiation regulation. KEGG analysis demonstrates that IVL mutations are associated with cAMP signaling pathway. IVL over-expression inhibits the proliferation and migration of esophageal cancer cells(P＜0.05). In addition, Q439H mutant IVL oligopeptides shows lower binding affinity to TGase1 than wild-type oligopeptides. Conclusions The seventh amino acid mutation in IVL repeat motif does weaken IVL binding to TGase1. This mutation may affect tumor-related biological processes and signaling pathways.

Key words: adenocarcinoma at esophagogastric junction, Involucrin, gene mutation

中图分类号:

吕雪, 李雪薇, 杨婷, 郑锦秀, 祝子鹤, 杨涛, 徐钧. 食管胃结合部腺癌生物信息学分析及Involucrin基因突变位点筛选[J]. 基础医学与临床, 2022, 42(2): 235-242.

LYU Xue, LI Xue-wei, YANG Ting, ZHENG Jin-xiu, ZHU Zi-he, YANG Tao, XU Jun. Bioinformatic analysis of adenocarcinoma at esophagogastric junction and mutation sites screening of Involucrin gene[J]. Basic & Clinical Medicine, 2022, 42(2): 235-242.

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食管胃结合部腺癌生物信息学分析及Involucrin基因突变位点筛选

Bioinformatic analysis of adenocarcinoma at esophagogastric junction and mutation sites screening of Involucrin gene

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