肿瘤相关基因ADAM15在结肠癌中一个非同义变异对细胞黏附及侵袭的影响  

A nonsynonymous single nucleotide variant of the tumor-related gene ADAM15 in colon cancer and its impact on cell adhesion phenotype

作　　者：王晶[1] 贺礼兵 张国燕 刘筱涵 程杉[1] Wang Jing;He Libing;Zhang Guoyan;Liu Xiaohan;Cheng Shan(Department of Medical Genetics and Developmental Biology,School of Basic Medical Sciences,Capital Medical University,Beijing 100069,China;Beijing No.11 High School,Beijing 100039,China)

机构地区：[1]首都医科大学基础医学院医学遗传学与发育生物学学系,北京100069 [2]北京市十一学校,北京100039

出　　处：《首都医科大学学报》2023年第6期1044-1052,共9页Journal of Capital Medical University

摘　　要：目的解析ADAM15基因单核苷酸变异(single nucleotide variant,SNV)在结肠癌肿瘤发生发展相关的细胞学过程中的具体功能作用。方法首先对具有侵袭表型的结肠癌患者癌组织及癌旁组织全外显子组测序,然后进行野生型/突变型ADAM15结肠癌HCT-8细胞系细胞功能实验,进而完成野生型/突变型ADAM15细胞系的转录组测序分析。结果采用全外显子组测序在预后不良的患者中检出ADAM15基因SNV rs6427128,提示可能影响结肠癌的进展。细胞功能实验显示,rs6427128损失了ADAM15所具有的上调细胞侵袭能力的作用,而使细胞黏附能力增加约26%。转录组测序分析显示,与野生型相比,rs6427128过表达细胞的差异表达基因富集在细胞黏附、DNA的复制与转录、炎症反应等多种细胞生物学过程,提示rs6427128变异可能通过促进肿瘤细胞的黏附参与调节肿瘤微环境的重塑,从而影响结肠癌的进展。结论结合适当的细胞模型和较为精细的分析方法,深入解析临床样本中所检出的一些高频非同义潜在功能性变异体,可以为肿瘤相关基因的结构功能关系提供有益的提示性实验室数据。Objective To elucidated the specific functional roles of the a disintegrin and metalloproteinase 15(ADAM15)gene single nucleotide variant(SNV)in the cellular processes associated with the development of colon cancer through combination of high throughput sequencing and suitable cellular models.Methods We initially performed whole exome sequencing on cancerous tissues and adjacent tissues from colon cancer patients exhibiting invasive phenotypes.Subsequently,functional experiments were conducted using wild-type/mutant ADAM15 colon cancer HCT-8 cell lines.This was followed by transcriptome sequencing analysis of the wild-type/mutant ADAM15 cell lines.Results The whole exome sequencing study identified a highly recurrent SNV,rs6427128,located at the ADAM15 gene,which demonstrated a significant association with adverse prognosis in patients,suggesting its potential influence on the progression of colon cancer.The functional assays utilizing cells overexpressing the variant gene,indicated that the expression product of this variant gene lost the effect of upregulating cell invasion capabilities while increased cell adhesion by approximately 26%compared to wild-type ADAM15 cells.To comprehensively investigate the mechanisms underlying the impact of rs6427128 variation on colon cancer cell phenotypes,we performed transcriptome sequencing analysis on HCT-8 cells transfected with the overexpressed variant protein.The results revealed differential expression genes were enriched in various cellular biological processes,including cell adhesion junction pathways,DNA replication and transcription,as well as inflammatory responses,in comparison to wild-type samples.Conclusion This study indicated that the rs6427128 variant may alter the tumor microenvironment by affecting tumor cell adhesion and contributed to extensive transcriptional changes in tumor cells,ultimately promoted the malignant phenotype of colon cancer.Although individual gene mutations in malignant tumors often do not act as primary causative factors,high-freque

关 键 词：结肠癌 ADAM15基因 细胞黏附 单核苷酸变异 肿瘤微环境 

分 类 号：R753.3[医药卫生—皮肤病学与性病学]