肠型胃癌的超级增强子重塑特征及上调迁移诱导蛋白CEMIP表达的作用研究  

Remodeling characteristics of super-enhancers in intestinal gastric cancer and their roles in upregulating CEMIP expression

作　　者：彭仕银 谭玉婷 薛锐 李先锋 王涛[2] 储召乐 刘碧颖 陈东风[2] 王斌 PENG Shiyin;TAN Yuting;XUE Rui;LI Xianfeng;WANG Tao;CHU Zhaole;LIU Biying;CHEN Dongfeng;WANG Bin(School of Medicine,Chongqing University,Chongqing;Department of Gastroenterology,Chongqing Key Laboratory of Precise Prevention and Treatment of Digestive Malignancies,Army Medical Center of PLA/Daping Hospital of Third Military Medical University,Chongqing,China)

机构地区：[1]重庆大学医学院,重庆 [2]陆军特色医学中心(第三军医大学大坪医院)消化内科,消化系统肿瘤精准防治重庆市重点实验室,重庆

出　　处：《陆军军医大学学报》2025年第5期396-406,共11页Journal of Army Medical University

基　　金：国家自然科学基金优秀青年科学项目(81822032);重庆市自然科学基金杰出青年项目(CSTC2019JCYJJQX0027)。

摘　　要：目的绘制肠型胃癌的超级增强子重塑图谱,揭示超级增强子的肿瘤生物学功能及可能激活的下游靶基因。方法收集陆军特色医学中心消化内科2022年1-12月收治行内镜检查、治疗的患者的31例正常胃黏膜组织、23例肠型胃癌组织及9例肠型胃癌类器官进行靶向组蛋白H3K27ac修饰的染色质靶向捕获(cleavage under targets and tagmentation,CUT&Tag)测序,基于生物信息学工具描绘肠型胃癌的超级增强子重塑特征并鉴定关键靶基因;利用CRISPRi技术干预超级增强子,Western blot检测靶基因的表达,并采用CCK-8法和Transwell迁移、侵袭实验检测对照组细胞及干预组细胞的增殖、迁移、侵袭能力。结果肠型胃癌组织与正常胃黏膜组织超级增强子信号差异明显(P<0.05),癌组织中的活性超级增强子可能参与免疫系统的负向调节、细胞黏附等生物学过程;在肿瘤细胞中表达上调的细胞迁移诱导蛋白(cell migration-inducing protein,CEMIP)受到超级增强子的调控,干预超级增强子后肿瘤细胞CEMIP蛋白表达量下降(P<0.05),细胞增殖率、相对迁移面积和相对侵袭面积也随之下降(P<0.05)。结论肠型胃癌发生了超级增强子重塑,其下游靶基因CEMIP可促进癌细胞生长、迁移和侵袭。Objective To map the super-enhancers remodeling of intestinal gastric cancer and reveal the tumor biological functions of the super-enhancers and the downstream target genes that may be activated.Methods A total of 31 normal gastric mucosal tissues,23 intestinal gastric cancer tissues and 9 intestinal gastric cancer organoids were collected from the Department of Gastroenterology of Army Medical Center of PLA from January to December 2022.Chromatin targeting histone H3K27ac modified chromatin targeting cleavage under targets and tagmentation(CUT&Tag)sequencing was conducted on above tissues.The remodeling profiles of super-enhancers in intestinal gastric cancer were analyzed and the key target genes were identified based on bioinformation tools.CRISPRi technology was used to intervene with the superenhancers,the expression of target genes was detected with Western blotting,and the proliferation,migration and invasion abilities were detected by CCK-8 assay and Transwell chambers in the control group and the intervention group.Results There was a significant difference in the signal of super-enhancers between intestinal gastric cancer tissues and normal gastric mucosal tissues(P<0.05),and the active super-enhancers in cancer tissues may be involved in biological processes such as negative regulation of the immune system and cell adhesion.The expression of up-regulated cell migration-inducing protein(CEMIP)in tumor cells was regulated by the super-enhancers,and intervening the super-enhancers down-regulated the expression of CEMIP(P<0.05),and inhibited the cell proliferation,invasion and migration abilities of tumor cells(P<0.05).Conclusion Super-enhancer remodeling is observed in intestinal gastric cancer,and they can up-regulate the expression of CEMIP gene and promote the growth,migration and invasion of cancer cells.

关 键 词：肠型胃癌 组蛋白H3K27ac 超级增强子 细胞迁移诱导蛋白 

分 类 号：R394.3[医药卫生—医学遗传学] R730.23[医药卫生—基础医学] R735.2