环状RNA_0023990/微小RNA-873-5p/ANXA2对甲状腺癌放射敏感性的影响及机制研究  

The mechanism of circ_0023990/miR-873-5p/ANXA2 axis regulating radiosensitivity and development of thyroid carcinoma

作　　者：李红光[1] 赵利红 卢昂 刘建波[3] 苏自杰 王晓博[4] 高永举[4] Li Hongguang;Zhao Lihong;Lu Ang;Liu Jianbo;Su Zijie;Wang Xiaobo;Gao Yongju(Department of Thyroid Surgery,Henan Provincial People′s Hospital,Zhengzhou,450003,China;Department of Disinfection Supply Center,Fuwai Central China Cardiovascular Hospital,Zhengzhou,450003,China;Department of Radiotherapy,Henan Provincial People′s Hospital,Zhengzhou,450003,China;Department of Nuclear Medicine,Henan Provincial People′s Hospital,Zhengzhou,450003,China)

机构地区：[1]河南省人民医院甲状腺外科,郑州450003 [2]阜外华中心血管病医院消毒供应中心,郑州450003 [3]河南省人民医院放疗科,郑州450003 [4]河南省人民医院核医学科,郑州450003

出　　处：《中华医学杂志》2021年第40期3329-3337,共9页National Medical Journal of China

基　　金：国家自然科学基金(81801762)。

摘　　要：目的探究circ_0023990对甲状腺癌细胞放射敏感性的影响及可能机制。方法采用逆转录聚合酶链反应(qRT-PCR)法检测2016年8月至2018年11月于河南省人民医院就诊的55例甲状腺癌患者[其中男12例,女43例,年龄(53.3±7.3)岁]癌组织及甲状腺癌细胞系(TPC-1、KTC-1、FTC-133和CAL-62)中circ_0023990的表达,分析癌组织中circ_0023990表达与患者临床病理特征的关系。甲状腺癌细胞TPC-1和KTC-1均分为circ_0023990小干拢RNA(sh-circ_0023990)组、无意义阴性序列(sh-NC)组、sh-circ_0023990+miR-873-5p抑制剂(anti-miR-873-5p)组、sh-circ_0023990+阴性对照序列(anti-miR-NC)组、miR-873-5p组、对照序列(miR-NC)组、miR-873-5p+pcDNA-ANXA2组和miR-873-5p+pcDNA组,克隆形成实验检测细胞放射敏感性;且各组细胞用4 Gy放射线照射后,蛋白质印迹法检测细胞中γH2AX蛋白表达。双荧光素酶报告基因实验验证circ_0023990与miR-873-5p及miR-873-5p与ANXA2的靶向关系。结果circ_0023990在甲状腺癌组织表达高于正常组织(2.15±0.09比0.97±0.05,P<0.05),且其表达与甲状腺癌患者肿瘤大小、淋巴结转移及TNM分期密切相关(P<0.05)。甲状腺癌TPC-1、KTC-1、FTC-133和CAL-62细胞中circ_0023990表达高于正常甲状腺细胞HTori-3(3.16±0.38、2.63±0.28、1.82±0.24、1.71±0.22比1.00±0.10,P均<0.05)。sh-circ_0023990组TPC-1和KTC-1细胞存活分数明显低于sh-NC组(P<0.05),增敏比分别为2.482、1.643;sh-circ_0023990+anti-miR-873-5p组TPC-1和KTC-1细胞存活分数高于sh-circ_0023990+anti-miR-NC组(P<0.05),增敏比分别为0.305、0.441。miR-873-5p组TPC-1、KTC-1细胞存活分数低于miR-NC组(P<0.05),增敏比分别为2.044、1.653;miR-873-5p+pcDNA-ANXA2组TPC-1、KTC-1细胞存活分数高于miR-873-5p+pcDNA组(P<0.05),增敏比分别为0.496、0.686。4 Gy+sh-circ_0023990组TPC-1和KTC-1细胞中γH2AX蛋白表达高于4 Gy+sh-NC组(2.68±0.27比1.87±0.25,2.46±0.19比1.77±0.14;P均<0.05),4Gy+sh-circ_0023990+anti-miR-873-5p组TPC-1和KObjective To explore the effect and possible mechanism of circ_0023990 on the radiosensitivity of thyroid cancer cells.Methods qRT-PCR was used to detect the expression of circ_0023990 in the cancer tissues of 55 patients with thyroid cancer and thyroid cancer cell lines(TPC-1,KTC-1,FTC-133 and CAL-62),and the relationship between the expression of circ_0023990 in cancer tissues and the clinical characteristics of the patients were analyzed.Thyroid cancer cells TPC-1 and KTC-1 were divided into sh-circ_0023990 group,sh-NC group,sh-circ_0023990+anti-miR-873-5p group,sh-circ_0023990+anti-miR-NC group,miR-873-5p group,miR-NC group,miR-873-5p+pcDNA-ANXA2 group and miR-873-5p+pcDNA group,and then clone formation experiment was used to detect cell radiosensitivity.After each group of cells was irradiated with 4Gy radiation,the expression of γH2AX protein in the cells was detected by Western Blot.The dual luciferase reporter gene experiment verified the targeting relationship between circ_0023990 and miR-873-5p or miR-873-5p and ANXA2.Results The expression of circ_0023990 in thyroid cancer tissues was higher than that in normal tissues(2.15±0.09 vs.0.97±0.05,P<0.05),and its expression was closely related to tumor size,lymph node metastasis and TNM staging of patients with thyroid cancer(P<0.05).The expression of circ_0023990 in thyroid cancer cell lines(TPC-1,KTC-1,FTC-133 and CAL-62)were higher than that of normal thyroid cells HTori-3(3.16±0.38,2.63±0.28,1.82±0.24,1.71±0.22 vs.1.00±0.10,all P<0.05).The survival scores of TPC-1 and KTC-1 cells in the sh-circ_0023990 group were significantly lower than those in the sh-NC group(P<0.05),and the sensitization ratios were 2.482,1.643;The survival scores of TPC-1 and KTC-1 cells in the sh-circ_0023990+anti-miR-873-5p group were higher than those in the sh-circ_0023990+anti-miR-NC group(P<0.05),and the sensitization ratios were 0.305,0.441,respectively.The survival scores of TPC-1 and KTC-1 cells in the miR-873-5p group were lower than those in the miR-NC group(P<0.0

关 键 词：甲状腺癌 circ_0023990 miR-873-5p ANXA2 放射敏感性 

分 类 号：R736.1[医药卫生—肿瘤]