长链非编码RNA LINC00261通过调控miR-324-3p/EST1促进胃癌进展  

Long non-coding RNA LINC 00261 promotes progression of gastric cancer by regulating miR-324-3 p/EST 1

作　　者：谢锐 尹源 滕俊 梁红亮 XIE Rui;YIN Yuan;TENG Jun;LIANG Hongliang(Department of Gastroenterology,363 Hospital,Chengdu 610041,China)

机构地区：[1]三六三医院消化内科,成都610041

出　　处：《中国免疫学杂志》2024年第10期2101-2107,共7页Chinese Journal of Immunology

摘　　要：目的:探讨长链非编码RNA LINC00261通过干扰miR-324-3p表达调控E26转录因子1(EST1)水平促进胃癌(GC)的进展。方法:收集2018年6月至2020年10月在三六三医院接受手术治疗的GC患者(n=80)的癌组织及其相应癌旁正常组织作为研究样本,qRT-PCR检测LINC00261和miR-324-3p表达水平。分析LINC00261与临床病理参数之间的相关性。将siRNA(si-LINC00261)、miRNA模拟物(miR-324-3p)和miRNA抑制剂(miR-324-3p in)及其相应对照转染至MGC-803和SGC-7901细胞;克隆形成试验评估细胞增殖能力;Transwell试验评估细胞迁移和侵袭能力;Western blot检测E-cadherin、N-cadherin和ETS1蛋白表达水平;肿瘤异种移植实验检测LINC00261在体内的肿瘤形成能力;荧光素酶报告、RNA沉淀分析LINC00261、miR-324-3p及EST1之间的作用关系。结果:与癌旁组织相比,GC组织中LINC00261表达明显上调,差异具有统计学意义(P<0.05)。LINC00261表达与患者TNM分期、组织分化程度淋巴结转移及微血管侵犯有关(P<0.05);通过数据库预测、萤火虫荧光素酶实验及RNA免疫沉淀结果显示,LINC00261和miR-324-3p存在靶向作用关系;GC组织中miR-324-3p水平明显低于癌旁正常组织(P<0.05);miR-324-3p水平与LINC00261表达呈负相关(P<0.05);与in NC+si-NC组相比,in NC+si-LINC00261组细胞增殖、迁移和侵袭能力及N-cadherin表达明显被抑制(P<0.05),E-cadherin表达明显升高(P<0.05);与in NC+si-LINC00261组相比,siLINC00261+miR-324-3p in组细胞增殖、迁移和侵袭能力及N-cadherin表达明显提高(P<0.05),E-cadherin表达明显降低(P<0.05),Targetscan预测及萤火虫荧光素酶实验表明ETS1是miR-324-3p的下游结合位点;转染miR-324-3p后,ETS1蛋白水平显著下调(P<0.05),但转染miR-324-3p in后,ETS1蛋白水平显著上调(P<0.05);在GC组织中ETS1表达明显高于癌旁正常组织(P<0.05);miR-324-3p水平与ETS1呈负相关(P<0.05);转染si-LINC00261的MGC-803细胞产生的肿瘤重量低于转染siNC的MGC-803细胞产生�Objective:To investigate the role of long non-coding RNA LINC 00261 in regulating E 26 transcription factor 1(EST 1)by interfering with the expression of miR-324-3 p in promoting the development of gastric cancer(GC).Methods:Cancer tis⁃sues and corresponding adjacent normal tissues of GC patients(n=80)who underwent surgical treatment in 363 Hospital from June 2018 to October 2020 were collected as research samples,and the expression levels of LINC 00261 and miR-324-3 p were detected by qRT-PCR.The correlation between LINC 00261 and clinicopathological parameters were analyzed.siRNA(si-LINC 00261),miRNA mimic(miR-324-3 p),miRNA inhibitor(miR-324-3 p in)and their corresponding controls were transfected into MGC-803 and SGC-7901 cells.Clonogenesis assay was used to evaluate cell proliferation.The Transwell assay assessed cell migration and invasion.The protein expression levels of E-cadherin,N-cadherin and ETS 1 were detected by Western blot.Tumor xenotransplantation assay was used to detect the tumorigenesis of LINC 00261 in vivo.Luciferase report and RNA precipitation were used to analyze the interaction between LINC 00261,miR-324-3 p and EST 1.Results:Compared with adjacent tissues,the expression of LINC 00261 in GC tissues was significantly up-regulated with statistical significance(P<0.05).The expression of LINC 00261 was correlated with TNM stage,tissue differentiation degree,lymph node metastasis and microvascular invasion(P<0.05).The database prediction,firefly luciferase assay and RNA immunoprecipitation results showed that LINC 00261 had a targeted relationship with miR-324-3 p.The level of miR-324-3 p in GC tissues was significantly lower than that in paracellular normal tissues(P<0.05).There was a negative correlation between miR-324-3 p level and LINC 00261 expression(P<0.05).Compared with in NC+si-NC group,the cell proliferation,migration and invasion ability and the expression of N-cadherin in in NC+si-LINC 00261 group were significantly inhibited(P<0.05),while the expression of E-cadherin was signi

关 键 词：胃癌 侵袭 增殖 长链非编码RNA LINC00261 E26转录因子1 

分 类 号：R735.2[医药卫生—肿瘤]