miRNA-210-3p在大肠癌细胞迁移侵袭中的促进作用及其靶基因的生物信息学分析  

Effect of miRNA-210-3p in Invasion of Colorectal Cancer Cells and Bioinformatic Analysis of Target Genes

作　　者：潘鑫[1] 李婉明[1] 于敏[1] 方瑾[1] Pan Xin;Li Wanming;Yu Min(Department of Cell Biology,Key Laboratory of Cell Biology,Ministry of Public Health,Key Laboratory of Medical Cell Biology,Ministry of Education,China Medical University,Liaoning 110122,China)

机构地区：[1]中国医科大学卫生部细胞生物学重点实验室、医学细胞生物学教育部重点实验室、生命科学院细胞生物学教研室,沈阳110122

出　　处：《医学研究杂志》2020年第9期51-56,共6页Journal of Medical Research

基　　金：国家自然科学基金资助项目(81672920,81502703);辽宁省沈阳市中青年科技创新人才支持计划项目(BC190261)。

摘　　要：目的探讨miR-210-3p在大肠癌细胞迁移、侵袭中的作用,并利用生物信息学方法预测其靶基因及信号通路。方法采用dbDEMC和OncomiR数据库分析miR-210-3p在大肠癌中的表达情况,通过荧光定量PCR法验证miR-210-3p在大肠癌细胞系中的表达;通过转染miRNA模拟物、抑制剂上调和下调大肠癌细胞内miR-210-3p的表达量,Transwell法检测转染后大肠癌细胞的迁移和侵袭能力变化;通过miRDB、TargetScan7.2、starBase和miRPathDB数据库预测miR-210-3p靶基因,利用DAVID数据库进行GO分析及信号通路的富集分析,利用GEPIA数据库分析关键基因在大肠癌组织中的表达。结果dbDEMC和OncomiR数据库显示miR-210-3p在大肠癌组织中高表达,并与肿瘤转移和患者预后相关。荧光定量PCR验证miR-210-3p在高转移性结肠癌细胞系HCT-116、LoVo中的表达量高于中转移性细胞系SW480和低转移性细胞系CL187、HCT-8。过表达miR-210-3p能明显增强SW480细胞迁移和侵袭能力;下调miR-210-3p的表达后,LoVo细胞迁移和侵袭能力则明显减弱。共预测出3035个靶基因,其功能主要富集在细胞迁移、黏附、蛋白质代谢等生物学过程以及癌症通路、HIF、Rap1等信号转导通路,GEPIA数据库验证显示RGMA在临床大肠癌组织中呈显著低表达,可能是miR-210-3p的靶基因。结论miR-210-3p可促进大肠癌细胞迁移和侵袭能力,有望成为大肠癌治疗的新型生物靶点。Objective To investigate the effect of miR-210-3p in colorectal cancer(CRC)cells migration and invasion and to analyze the enriched signaling pathways of target genes by bioinformatic methods.Methods The expression of miR-210-3p in colorectal cancer was analyzed by dbDEMC and OncomiR database.Real-time quantitative PCR was used to detecte the expression of miR-210-3p in five types of CRC cells(CL187,HCT-8,SW480,HCT-116,LoVo).miRNA mimics or inhibitor was transfected in CRC cells to up or down regulate the expression of miR-210-3p.Transwell assays were used to evaluated migration and invasion of CRC cells after transfection.Target gengs were pridected by miRDB,TargetScan7.2,starBase and miRPathDB.Gene ontology analysis and signal pathway enrichment were performed by DAVID.Results dbDEMC and OncomiR database showed that miR-210-3p was highly expressed in colorectal cancer,which was related to tumor metastasis and prognosis.Real-time quantitative PCR tested expression of miR-210-3p in highly invasiive cell lines(HCT-116 and LoVo)was significantly higher than that in moderate invasive cell lines SW480.And low invasive cell lines(CL187,HCT-8)revealed the lowest expression.Overexpression of miR-210-3p significantly enhanced SW480 cell migration and invasion,while LoVo cell migration and invasion was decreased after knockdown miR-210-3p.A total of 3035 target genes were predicted and the functions were mainly enriched in biological processes such as cell migration,adhesion,protein metabolism.The enriched pathways include cancer pathway,HIF signaling pathway and Rap1 signaling pathway.RGMA is significantly down regulated in clinical CRC tissues and may be the target gene of miR-210-3p.Conclusion miR-210-3p can improve the migration and invasion abilities of colorectal cancer cells,which indicates that it might be a new biological target for colorectal cancer treatment.

关 键 词：miR-210-3p 大肠癌 侵袭 生物信息学 靶基因 

分 类 号：R34[医药卫生—基础医学]