TNRC6C-AS1和miR-129-5p对甲状腺癌细胞增殖、迁移和侵袭的影响  

Effect of TNRC6C-AS1 and miR-129-5p on proliferation,migration and invasion of thyroid cancer cells

作　　者：李化静[1] 任婉丽[1] 郝润梅 汪世洋 白艳霞[1] 赵谦[1] LI Huajing;REN Wanli;HAO Runmei;WANG Shiyang;BAI Yanxia;ZHAO Qian(Department of Otolaryngology-Head and Neck Surgery,First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,China)

机构地区：[1]西安交通大学第一附属医院耳鼻咽喉头颈外科,西安710061

出　　处：《山西医科大学学报》2021年第7期831-839,共9页Journal of Shanxi Medical University

基　　金：陕西省科技计划-重点研发计划项目(2020-SF-A20);陕西省自然科学基础研究计划项目(2020JM-382)。

摘　　要：目的探究lncRNA TNRC6C-AS1和miR-129-5p对甲状腺癌(TC)细胞的增殖、迁移和侵袭的影响。方法收集2018年1月至2020年6月期间本院收治的40例甲状腺癌患者的甲状腺肿瘤组织和邻近正常对照甲状腺组织样品。将人甲状腺癌细胞FTC-133分为空载体阴性对照组(空载体组)、miR-129-5p模拟物组(模拟物组)、miR-129-5p抑制剂组(抑制剂组)、sh-TNRC6C-AS1+miR-129-5p抑制剂组(sh-AS1+抑制剂组)、sh-Unc5b组和sh-TNRC6C-AS1(sh-AS1)组。采用qRT-PCR检测甲状腺癌细胞系(NPA87、KAT-5和FTC-133)和正常人甲状腺细胞系HTori3及甲状腺肿瘤组织中TNRC6C-AS1、miR-129-5p和Unc5b的表达水平。采用Western blotting检测TNRC6C-AS1、miR-129-5p和Unc5b的表达水平;CCK-8法检测细胞增殖;Transwell法检测细胞迁移和侵袭。通过生物信息学和双荧光素酶报告分析预测TNRC6C-AS1、miR-129-5P和Unc5b之间的相互作用。结果与甲状腺癌旁临近正常组织和正常细胞HTori3相比,TC组织和细胞系(NPA87、KAT-5和FTC-133)中的TNRC6C-AS1和Unc5b的表达水平显著升高,miR-129-5p的表达水平显著降低(均P<0.05)。在FTC-133细胞中,与空载体组相比,sh-AS1组的TNRC6C-AS1和Unc5b相对表达水平显著降低,miR-129-5p相对表达水平显著升高(均P<0.01);模拟物组中Unc5b的相对表达水平显著降低,miR-129-5p相对表达水平显著升高(均P<0.01);抑制剂组中miR-129-5p的相对表达水平显著降低(P<0.01);sh-AS1+抑制剂组中miR-129-5p的相对表达量无显著变化(P>0.05)。与空载体组相比,sh-AS1组和sh-Unc5b组FTC-133细胞的增殖、迁移和侵袭能力显著降低(P<0.01);模拟物组FTC-133细胞的增殖、迁移和侵袭能力显著降低(P<0.01);抑制剂组FTC-133细胞的增殖、迁移和侵袭能力显著升高(P<0.01);sh-AS1+抑制剂组FTC-133细胞的增殖、迁移和侵袭能力无显著变化(P>0.05)。双荧光素酶报告结果显示,miR-129-5p与AS1-wt的3′-UTR结合,Unc5b为miR-129-5p的靶基因。结论TNRC6C-ASObjective To explore the effects of lncRNA TNRC6C-AS1 and miR-129-5p on the proliferation,migration and invasion of thyroid cancer(TC)cells.Methods Forty thyroid tumor tissue samples and adjacent normal control thyroid tissue samples were collected from thyroid cancer patients admitted to our hospital from January 2018 to June 2020.The human thyroid cancer cells FTC-133 were divided into empty negative control group(empty vector group),miR-129-5p mimic group(mimic group),miR-129-5p inhibitor group(inhibitor group),sh-TNRC6C-AS1+miR-129-5p inhibitor group(sh-AS1+inhibitor group),sh-Unc5b group and sh-TNRC6C-AS1 group(sh-AS1 group).The expression levels of TNRC6C-AS1,miR-129-5p and Unc5b in thyroid cancer cell lines(NPA87,KAT-5 and FTC-133),normal human thyroid cell line HTori3 and thyroid cancer tissues were detected by qRT-PCR.The expression levels of TNRC6C-AS1,miR-129-5p and Unc5b were detected by Western blotting,the cell proliferation was detected by CCK-8 method,and the cell migration and invasion abilities were detected by Transwell method.The interaction between TNRC6C-AS1,miR-129-5p and Unc5b was predicted by bioinformatics and double luciferase report analysis.Results Compared with adjacent normal tissues and normal cells HTori3,the expression levels of lncRNA TNRC6C-AS1 and Unc5b in TC tissues and cells(NPA87,KAT-5 and FTC-133)were significantly increased,while the expression of miR-129-5p was significantly decreased(all P<0.05).In FTC-133 cells,compared with empty vector group,the expression levels of TNRC6C-AS1 and Unc5b were significantly decreased in sh-AS1 group,while the relative expression of miR-129-5p was significantly increased(all P<0.01);the relative expression of Unc5b was significantly decreased in mimic group,while the relative expression of miR-129-5p was significantly increased(all P<0.01);the relative expression of miR-129-5p was significantly decreased in inhibitor group(P<0.01).Compared with empty vector group,the relative expression of miR-129-5p showed no significant change in sh-A

关 键 词：miR-129-5p 甲状腺癌 TNRC6C-AS1 Unc5b 

分 类 号：R736.1[医药卫生—肿瘤]