lncRNA CTD-2182N23.1在甲状腺乳头状癌组织中的表达及其对癌细胞恶性生物学行为的影响  

The expression of lncRNA CTD-2182N23.1 in papillary thyroid carcinoma tissues and its influence on the malignant biological behavior of papillary thyroid carcinoma cells

作　　者：黎思毅 肖雄升 陈卓婷 杨雪[2] 郑昌蔚 张智 LI Siyi;XIAO Xiongsheng;CHEN Zhuoting;YANG Xue;ZHENG Changwei;ZHANG Zhi(Department of Vascular and Thyroid Surgery,Affiliated Hospital of Guangdong Medical University,Guangdong Zhanjiang 524000,China;Department of Oncology,Affiliated Hospital of Qingdao University,Shandong Qingdao 266003,China)

机构地区：[1]广东医科大学附属医院血管甲状腺外科,广东湛江524000 [2]青岛大学附属医院肿瘤科,山东青岛266003

出　　处：《现代肿瘤医学》2024年第1期1-7,共7页Journal of Modern Oncology

基　　金：国家自然科学基金(编号:81801734)。

摘　　要：目的:分析长链非编码(long non-coding RNA,lncRNA)CTD-2182N23.1在甲状腺乳头状癌组织中的表达水平与患者临床病理特征的关系,探究CTD-2182N23.1通过靶向miR-200c-3p对甲状腺乳头状癌细胞恶性生物学行为的影响。方法:采用TCGA数据库分析CTD-2182N23.1在甲状腺乳头状癌组织中的表达及其与患者临床分期、预后的关系。收集2019年05月至2022年03月本院手术切除的50例甲状腺乳头状癌患者癌组织和癌旁组织,采用实时定量聚合酶链反应(qPCR)检测甲状腺乳头状癌组织中CTD-2182N23.1的表达,分析其与甲状腺乳头状癌患者临床病理特征的关系。qPCR检测人正常甲状腺滤泡上皮细胞Nthy-ori3-1和人甲状腺乳头状癌细胞系TPC-1、MDA-T32、TT、SW579、B-CPAP中CTD-2182N23.1的表达。体外培养MDA-T32细胞,实验分为NC组和CTD-2182N23.1组。细胞克隆形成实验和Transwell实验分别检测转染后MDA-T32细胞增殖能力和侵袭能力。双荧光素酶基因报告实验验证CTD-2182N23.1与miR-200c-3p之间的靶向关系。qPCR检测转染后MDA-T32细胞中miR-200c-3p和ATP2A2 mRNA的表达。采用TCGA数据库分析CTD-2182N23.1与ATP2A2 mRNA表达的相关性。Western blot法分别检测ATP2A2、CDK1、Cyclin B、Twist、Slug蛋白水平。结果:TCGA数据库显示甲状腺乳头状癌组织中CTD-2182N23.1呈低表达(P<0.01),其与患者临床分期、无病生存期均呈正相关(均P<0.05)。qPCR显示甲状腺乳头状癌组织和细胞系中CTD-2182N23.1均呈低表达(均P<0.05)。CTD-2182N23.1表达水平与肿瘤分期、淋巴结转移有关联(均P<0.01)。上调CTD-2182N23.1后MDA-T32细胞克隆形成数目和细胞侵袭数目均明显减少(均P<0.01)。CTD-2182N23.1能够靶向且负调控miR-200c-3p的表达(P<0.01)。上调CTD-2182N23.1后MDA-T32细胞中ATP2A2基因表达显著升高(P<0.01)。CTD-2182N23.1与ATP2A2 mRNA表达呈正相关。上调CTD-2182N23.1后MDA-T32细胞中CDK1、Cyclin B、Twist、Slug蛋白水平均显著降�Objective:To analyze the relationship between the expression level of long non-coding RNA(lncRNA) CTD-2182N23.1 in papillary thyroid carcinoma tissues and the clinicopathological characteristics of patients.To explore the effect of CTD-2182N23.1 on the malignant biological behavior of papillary thyroid carcinoma cells by targeting miR-200c-3p.Methods:The TCGA database was used to analyze the expression of CTD-2182N23.1 in papillary thyroid carcinoma tissues and its relationship with the clinical stage and prognosis of patients.From May2019 to March 2022,the cancer tissues and paracancerous tissues of 50 patients with papillary thyroid carcinoma who underwent surgical resection in our hospital were collected,and real-time quantitative polymerase chain reaction(qPCR) was used to detect the expression of CTD-2182N23.1 in papillary thyroid carcinoma tissues.The relationship between its expression and the clinicopathological features of patients with papillary thyroid carcinoma was analyzed.The expression of CTD-2182N23.1 in human normal thyroid follicular epithelial cells Nthy-ori3-1 and human papillary thyroid carcinoma cell lines TPC-1,MDA-T32,TT,SW579,and B-CPAP was detected by qPCR.MDA-T32 cells were cultured in vitro and divided into NC group and CTD-2182N23.1 group.Cell clone formation assay and Transwell assay were used to detect the proliferation and invasion abilities of transfected MDA-T32 cells,respectively.The dual luciferase gene reporter assay verified the targeting relationship between CTD-2182N23.1 and miR-200c-3p.The expression of miR-200c-3p and ATP2A2 mRNA in MDA-T32 cells after transfection was detected by qPCR.The correlation between CTD-2182N23.1 and ATP2A2 mRNA expression was analyzed using TCGA database.The protein levels of ATP2A2,CDK1,Cyclin B,Twist and Slug were detected by Western blot respectively.Results:The TCGA database showed that the expression of CTD-2182N23.1 in papillary thyroid carcinoma tissues was low(P< 0.01),which was positively correlated with the clinical stage and disease-

关 键 词：长链非编码RNA 甲状腺乳头状癌 miR-200c-3p ATP2A2 增殖 侵袭 

分 类 号：R736.1[医药卫生—肿瘤]