特异性核基质结合区结合蛋白1和微小RNA-495-3P在甲状腺乳头状癌侵袭和转移中的作用  

Roles of special adenine-thymine-rich-binding rich sequence binding protein-1 and microRNA-495-3P in invasion and metastasis of papillary thyroid carcinoma

作　　者：林钦 张丽婷 吴文艺[1] 林建清[1] 史海鸿[1] 吴鑫泉 余艺煌 丁明骥 黄种心[3] 邱建龙 Lin Qin;Zhang Liting;Wu Wenyi;Lin Jianqing;Shi Haihong;Wu Xinquan;Yu Yihuang;Ding Mingji;Huang Zhongxin;Qiu Jianlong(Department of Thyroid and Breast Surgery,the Second Affiliated Hospital of Fujian Medical University,Quanzhou 362000,China;Department of Endocrinology,the 910th Hospital of the People′s Liberation Army,Quanzhou 362000,China;Department of Pathology,the Second Affiliated Hospital of Fujian Medical University,Quanzhou 362000,China;Department of Pathology,the 910th Hospital of the People′s Liberation Army,Quanzhou 362000,China)

机构地区：[1]福建医科大学附属第二医院甲状腺乳腺外科,泉州362000 [2]解放军第九一〇医院内分泌科,泉州362000 [3]福建医科大学附属第二医院病理科,泉州362000 [4]解放军第九一〇医院病理科,泉州362000

出　　处：《中华实验外科杂志》2021年第1期139-143,共5页Chinese Journal of Experimental Surgery

基　　金：泉州市科技局高层次人才创业创新项目(2017Z017);泉州市科技计划项目(2018N002S、2018Z145);福建医科大学科研苗圃基金项目(2016MP11)。

摘　　要：目的探讨特异性核基质结合区结合蛋白1(SATB1)和微小RNA(microRNA,miR)-495-3P在甲状腺乳头状癌(PTC)侵袭和转移中的作用。方法2019年9月至2020年4月,福建医科大学附属第二医院甲状腺乳腺外科采集肿瘤标本,将取得的40对PTC组织作为实验组,与之对应40对癌旁正常组织作为对照组。采用实时定量反转录聚合酶链反应(RT-qPCR)检测40对PTC组织及癌旁正常组织中SATB1和miR-495-3p的表达水平,用蛋白质印迹法(Western blot)检测16对PTC组织及癌旁正常组织中SATB1蛋白的表达水平。用si-SATB1转染人甲状腺乳头状癌细胞(TPC)-1后,用RT-qPCR、Western blot检测TPC-1中SATB1、miR-495-3p的表达水平,并用Pearson分析法进行相关性分析,转染si-NC的TPC-1为阴性对照组,转染si-SATB1的TPC-1为实验组。分别用细胞计数试剂盒(CCK-8)法、流式细胞计数、Trsnawell法检测敲低SATB1的表达对TPC-1增殖、凋亡、周期、侵袭、迁移能力的影响。两样本比较采用t检验。结果与癌旁正常组织比较,SATB1 mRNA和SATB1蛋白在PTC组织中呈高表达(1.27±0.14比0.86±0.23,t=8.484,P<0.01;0.94±0.10比0.37±0.15,t=11.890,P<0.01),差异有统计学意义,miR-495-3p呈低表达(0.78±0.11比1.37±0.64,t=5.741,P<0.01),差异有统计学意义。SATB1和miR-495-3p的异常表达均与PTC淋巴结转移明显相关(1.34±0.14,t=2.576,P<0.05;0.74±0.07,t=2.187,P<0.05)。PTC中SATB1和miR-495-3p的表达水平呈负相关(r=-0.497,P<0.01)。干扰TPC-1中SATB1的表达会导致miR-495-3p的表达水平高于阴性对照组(8.59±0.16比1.01±0.02,t=81.420,P<0.01),并且抑制细胞的增殖能力(0.39±0.01、0.52±0.01、0.58±0.03比0.43±0.01、0.60±0.01、0.72±0.01,t=4.899、9.798、7.668,P<0.01)、促进细胞凋亡[(42.8±2.1)%比(7.6±0.7)%,t=27.540,P<0.01]、减弱细胞侵袭(75.33±3.51比206.33±5.51,t=34.730,P<0.01)、迁移(202.00±7.81比528.33±5.03,t=60.840,P<0.01)能力并发生G2/M周期阻滞[(36.7±1.2)%比(15.6±0.9)%,t=24.360,P<0.01]Objective To study the role of special adenine-thymine-rich-binding rich special AT-rich-binding protein 1(SATB1)and microRNA(miRNA,miR)-495-3P in the invasion and metastasis of papillary thyroid carcinoma(PTC).Methods All specimens were collected from department of Thyroid and Breast Surgery,Second Affiliated Hospital of Fujian Medical University from September 2019 to April 2020,including 40 cases of PTC and 40 cases of corresponding normal thyroid tissues.There were 10 males and 30 females,aged 18-68 years,with a median age of 43 years.Quantitative real-time polymerase chain reaction(RT-qPCR)was used to detect the expression levels of SATB1 and miR-495-3p in 40 pairs of PTC tissues and normal tissues adjacent to cancer.Western blotting was used to detect the expression levels of SATB1 protein in 16 pairs of PTC tissues and normal tissues adjacent to cancer.After transfecting Human thyroid papillary carcinoma cells(TPC-1)with si-SATB1,RT-qPCR and Western blotting were used to detect the expression levels of SATB1 and miR-495-3p in TPC-1,and Pearson analysis method was used for correlation analysis.Methyl thiazol tetrazolium(MTT)method,flow cytometry and Trsnawell method were used to detect the effect of knocking down SATB1 expression on TPC-1 proliferation,apoptosis,cycle,invasion and migration.Results Compared with normal tissues adjacent to cancer,SATB1 mRNA and SATB1 proteins were highly expressed in PTC tissues(1.27±0.14 vs.0.86±0.23,t=8.484,P<0.01 and 0.94±0.10 vs.0.37±0.15,t=11.890,P<0.01),and miR-495-3p was down-regulated(0.78±0.11 vs.1.37±0.64,t=5.741,P<0.01).The abnormal expression of SATB1 and miR-495-3p was significantly correlated to PTC lymph node metastasis(1.34±0.14,t=2.576,P<0.05 and 0.74±0.07,t=2.187,P<0.05).The expression levels of SATB1 and miR-495-3p in PTC were negatively correlated(r=-0.497,P<0.01).Interfering with the expression of SATB1 in TPC-1 could increase the expression level of miR-495-3p(8.59±0.16 vs.1.01±0.02,t=81.420,P<0.01),and inhibit cell proliferation(0.39±0.01,0.

关 键 词：甲状腺癌 微小RNA 侵袭 凋亡 

分 类 号：R736.1[医药卫生—肿瘤]