LncRNA SNHG5/miR-26a-5p/MTDH信号轴促进结直肠癌转移的机制研究  

A study on mechanism of lncRNA-mediated SNHG5/miR-26a-5p/MTDH signal axis promoting metastasis of colorectal cancer

作　　者：冶俊玲[1] 郑小影 郭新建[1] 陈瑞慧 杨柳 苟笑丹[1] 蒋汉梅[2] YE Junling;ZHENG Xiaoying;GUO Xinjian;CHEN Ruihui;YANG Liu;GOU Xiaodan;JIANG Hanmei(Department of Pathology,Affiliated Hospital of Qinghai University,Xining 810001,Qinghai Provinve,China;Department of Gastroenterology,Affiliated Hospital of Qinghai University,Xining 810001,Qinghai Provinve,China)

机构地区：[1]青海大学附属医院病理科,青海西宁810001 [2]青海大学附属医院消化内科,青海西宁810001

出　　处：《中国癌症杂志》2023年第7期673-685,共13页China Oncology

基　　金：青海省卫生健康委员会一般指导性课题(2020-wjzdx-46)。

摘　　要：背景与目的:长链非编码RNA小核仁RNA宿主基因5(long non-coding RNA small nucleolar RNA host gene 5,lncRNA SNHG5)在多种癌症中发挥促癌作用,但其对结直肠癌(colorectal cancer,CRC)的影响和调节机制尚不清楚。本研究旨在探究lncRNA SNHG5/miR-26a-5p/异粘蛋白(metadherin,MTDH)信号轴促进CRC转移的机制。方法:分析癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库中的数据,探索CRC中异常表达的lncRNA并进行生存分析。收集2020年10月—2021年10月经手术切除的100例CRC、癌旁组织样本及患者的完整临床资料,采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR)检测lncRNA SNHG5、miR-26a-5p表达水平,采用免疫组织化学法检测MTDH的表达水平,分析lncRNA SNHG5在CRC中的相对表达水平与临床病理学特征及生存期的关系。通过细胞计数试剂盒-8(cell counting kit-8,CCK-8)检测、克隆形成、划痕、transwell实验及体内异种移植实验检测lncRNA SNHG5对CRC细胞增殖、迁移及侵袭的影响,通过蛋白质印迹法(Western blot)和免疫组织化学检测CRC细胞转移、上皮-间充质转化相关分子表达水平与lncRNA SNHG5表达水平的关系。通过RNA下拉实验、双荧光素酶报告基因检测、RNA免疫共沉淀实验探究SNHG5与miR-26a-5p、MTDH与miR-26a-5p在物理上的相互作用,通过CCK-8、EdU、transwell实验验证三者在功能上的相互关系,采用Western blot检测分析SNHG5、miR-26a-5p、MTDH表达对迁移、侵袭相关分子的影响。结果:TCGA数据库分析结果显示,lncRNA SNHG5在CRC中显著上调。RTFQ-PCR和免疫组织化学检测结果显示,CRC组织中lncRNA SNHG5、MTDH水平显著上调(P<0.05),miR-26a-5p水平下降(P<0.05),SNHG5高表达的样本中MTDH水平也较高。浆膜及浆膜外浸润、远处转移、淋巴结转移、TNMⅢ期的CRC组织lncRNA SNHG5表达量高于浆膜下浸润、无远处转移、无淋巴结转移、TNMⅠ~�Background and purpose:Long non-coding RNA small nucleolar RNA host gene 5(lncRNA SNHG5)plays a cancer-promoting role in many cancers,however its effect on colorectal cancer(CRC)and its regulatory mechanism are not clear.This study aimed to explore the mechanism of lncRNA SNHG5/miR-26a-5p/metadherin(MTDH)signal axis promoting metastasis of CRC.Methods:The data of The Cancer Genome Atlas(TCGA)database was analyzed,the abnormal expression of lncRNA in CRC was explored and analyzed the survival.Samples of CRC,paracancerous tissues and complete clinical data of patients who underwent surgical resection from October 2020 to October 2021 were collected.The expression levels of SNHG5 and miR-26a-5p in lncRNA were detected by real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR),and the expression level of MTDH was detected by immunohistochemistry.The relationship between the relative expression level of lncRNA SNHG5 in CRC and clinicopathological features and survival time was analyzed.The effects of lncRNA SNHG5 on the proliferation,migration and invasion of CRC cells were detected by cell counting kit-8(CCK-8),clone formation,scratching assays,transwell test and in vivo xenotransplantation.The relationship between CRC cell metastasis,the expression level of epithelial-mesenchymal transition related molecules and lncRNA SNHG5 expression level by Western blot and immunohistochemical detection were explored.The physical interaction between SNHG5 and miR-26a-5p,MTDH and miR-26a-5p was studied by RNA pull-down test,double luciferase reporter gene detection and RNA co-immunoprecipitation.The functional relationship among the three was verified by CCK-8,EdU and transwell experiments.The effect of SNHG5,miR-26a-5p and MTDH expression on migration and invasion related molecules was analyzed by Western blot.Results:The results of TCGA database analysis showed that lncRNA SNHG5 was significantly upregulated in CRC.The results of RTFQ-PCR and immunohistochemistry showed that the levels of lncRNA SNHG5 and MTDH i

关 键 词：长链非编码RNA小核仁RNA宿主基因5 miR-26a-5p 异黏蛋白 结直肠癌 转移 

分 类 号：R735.3[医药卫生—肿瘤]