miR-135b-3p调控XAF1表达影响甲状腺癌细胞的迁移、侵袭和放射增敏性  

miR-135b-3p regulates XAF1 expression and affects migration,invasion and radio sensitization of thyroid cancer cells

作　　者：曹邓晗[1] 陈奎生[2] CAO Deng-han;CHEN Kui-sheng(Department of Endocrinology,Zhengzhou First People’s Hospital,Zhengzhou 450004;Department of Pathology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州市第一人民医院内分泌科,河南郑州450004 [2]郑州大学第一附属医院病理科,河南郑州450052

出　　处：《基础医学与临床》2020年第6期790-796,共7页Basic and Clinical Medicine

基　　金：河南省医学科技攻关计划(201503221)。

摘　　要：目的探讨miR-135b-3p对甲状腺癌细胞迁移、侵袭和放射敏感性的影响以及其可能的调控机制。方法培养正常甲状腺细胞Nthy-ori 3-1、甲状腺癌细胞K1和TPC-1,RT-qPCR检测细胞中miR-135b-3p表达,Western blot检测细胞中X染色体连锁凋亡抑制蛋白相关因子1(XAF1)表达。转染anti-miR-135b-3p至TPC-1细胞抑制miR-135b-3p表达,Transwell、Western blot及克隆形成实验分别检测抑制miR-135b-3p表达对TPC-1细胞迁移和侵袭、E-cadherin和MMP-2蛋白表达及对TPC-1细胞放射敏感性的影响。双荧光素酶报告基因实验验证miR-135b-3p与XAF1之间的靶向调控关系。结果与Nthy-ori 3-1细胞相比,K1和TPC-1细胞中miR-135b-3p表达水平显著升高(P<0.05),XAF1蛋白表达水平显著降低(P<0.05)。抑制miR-135b-3p表达可抑制TPC-1细胞的迁移和侵袭能力(P<0.05),促进TPC-1细胞E-cadherin蛋白表达(P<0.05),抑制MMP-2蛋白表达(P<0.05),增强TPC-1细胞的放射敏感性(P<0.05)。miR-135b-3p靶向负调控XAF1表达。抑制XAF1表达可降低抑制miR-135b-3p表达对TPC-1细胞迁移、侵袭及放射敏感性的影响。结论miR-135b-3p通过下调XAF1表达促进甲状腺癌细胞的迁移和侵袭能力,并降低其放射增敏性,是甲状腺癌的潜在治疗靶点。Objective To investigate the effects of miR-135 b-3 p on migration,invasion and radio sensitivity of thyroid cancer cells and its possible regulatory mechanisms.Methods Normal thyroid cells Nthy-ori 3-1,thyroid cancer cells K1 and TPC-1 were cultured.The expression of miR-135 b-3 p was detected by RT-qPCR.The expression of XAF1 protein was detected by Western blot.Anti-miR-135 b-3 p was transfected to TPC-1 cells to inhibit the expression of miR-135 b-3 p.Transwell,Western blot and cloning formation assay were used to detect the effect of inhibition of miR-135 b-3 p expression on migration and invasion of TPC-1 cells,and the expression of E-cadherin and MMP-2 protein and radio sensitivity,respectively.The dual luciferase reporter gene assay verified the targetedregulatory relationship between miR-135 b-3 p and XAF1.Results Compared with Nthy-ori 3-1 cells,the expression of miR-135 b-3 p was significantly increased in K1 and TPC-1 cells(P<0.05),and the expression of XAF1 protein was significantly decreased(P<0.05).Inhibition of miR-135 b-3 p expression could inhibit the migration and invasion of TPC-1 cells(P<0.05),promote the expression of E-cadherin protein in TPC-1 cells(P<0.05),inhibit the expression of MMP-2 protein(P<0.05),and enhance the radio sensitivity of TPC-1 cells(P<0.05).miR-135 b-3 p targeted negative regulation of XAF1 expression.Inhibition of XAF1 expression partially reduced the effect of inhibiting miR-135 b-3 p expression on TPC-1 cell migration and invasion as well as the enhanced radio sensitivity.Conclusions R-135 b-3 p promotes the migration and invasion of thyroid cancer cells by down-regulating the expression of XAF1,and reduces its radio sensitization.It is a potential therapeutic target for thyroid cancer.

关 键 词：甲状腺癌 miR-135b-3p XAF1 迁移 放射增敏 

分 类 号：R736.1[医药卫生—肿瘤]