环状RNA circGPX1通过调节miRNA-150表达影响甲状腺癌细胞增殖和侵袭  

Circular RNA circGPX1 affects the proliferation and invasion of thyroid cancer cells by regulating the expression of miRNA-150

作　　者：孙崇镨 唐远怀 柴豆豆 陈伟[2] 孙超[1] Sun Chongpu;Tang Yuanhuai;Chai Doudou;Chen Wei;Sun Chao(Department of Thyroid Surgery,Weihai Municipal Hospital,Weihai 264200,China;Department of Oncology,the Second Affiliated Hospital of Nanjing Medical University,Nanjing 210003,China)

机构地区：[1]威海市立医院甲状腺外科,威海264200 [2]南京医科大学第二附属医院肿瘤科,南京210003

出　　处：《肿瘤研究与临床》2024年第4期255-261,共7页Cancer Research and Clinic

基　　金：国家自然科学基金(81902725)。

摘　　要：目的探讨环状RNA(circRNA)circGPX1在甲状腺癌组织中的表达及体外circGPX1对甲状腺癌细胞增殖和侵袭的影响和分子机制。方法基于GeneCards数据库分析circGPX1在甲状腺癌组织和癌旁组织中转录水平表达差异,比较不同临床分期患者circGPX1表达水平。采用实时荧光定量聚合酶链反应(qRT-PCR)法检测circGPX1在人正常甲状腺上皮细胞株Nthy-ori3-1和甲状腺癌细胞株SW579、TPC-1、B-CPAP、TT中的表达。将circGPX1表达水平最高的TPC-1细胞转染载有干扰circGPX1小干扰RNA(siRNA)序列的质粒和载有阴性对照siRNA序列的质粒,分别为si-circGPX1组和si-NC组。采用CCK-8法和Transwell实验分别检测两组TPC-1细胞增殖(以吸光度值为细胞增殖能力)和侵袭能力。采用CircAtlas软件预测circGPX1与miRNA-150(miR-150)互补,应用双荧光素酶报告基因实验验证二者的靶向关系。qRT-PCR法检测两组TPC-1细胞中miR-150表达。蛋白质印迹法检测两组TPC-1细胞中Wnt/β-连接素(catenin)信号通路β-catenin、c-myc、p-GSK3β、细胞周期蛋白D蛋白表达。结果GeneCards数据库中,与癌旁组织相比,甲状腺癌组织中circGPX1转录水平高表达,差异有统计学意义(P<0.01);甲状腺癌患者临床分期越高,circGPX1相对表达量越高,差异有统计学意义(P<0.01)。各甲状腺癌细胞株中circGPX1转录水平相对表达量均高于Nthy-ori3-1细胞,差异均有统计学意义(均P<0.01)。si-circGPX1组TPC-1细胞中circGPX1相对表达量较si-NC组低(1.2±0.4比6.1±0.5),差异有统计学意义(t=8.48,P=0.001)。铺板后36、48、60、72 h,si-circGPX1组TPC-1细胞的增殖能力均低于si-NC组,差异均有统计学意义(均P<0.01)。培养27 h,si-circGPX1组侵袭的TPC-1细胞数少于si-NC组[(40±6)个比(96±11)个],差异有统计学意义(t=4.30,P=0.005)。双荧光素酶报告基因实验显示,circGPX1-野生型与miR-150共转染的TPC-1细胞相对荧光素酶活性低于circGPX1-野生型与miR-150阴性对照�Objective To explore the expression of circular RNA(circRNA)circGPX1 in thyroid cancer tissues,and the effect of circGPX1 on the proliferation and invasion of thyroid cancer cells in vitro as well as its molecular mechanism.Methods Based on the GeneCards database,the transcriptional level differences of circGPX1 in thyroid cancer tissues and paracancerous tissues were analyzed,and the expression levels of circGPX1 in patients with different clinical stages were compared.Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of circGPX1 in human normal thyroid follicular epithelial cells Nthy-ori3-1 and thyroid cancer cell lines(SW579,TPC-1,B-CPAP,TT).TPC-1 cells with the highest expression level of circGPX1 were divided into si-circGPX1 group and si-NC group,which were transfected with plasmid carrying circGPX1 interfered by small interference RNA(Si-RNA)sequence and plasmid carrying negative control si-RNA sequence,respectively.CCK-8 assay and Transwell assay were used to detect the proliferation and invasion ability of TPC-1 cells in the above 2 groups(the absorbance value was used as the cell proliferation ability).CircAtlas software was used to predict that the circGPX1 was complementary to miRNA-150(miR-150).The dual luciferase gene reporter assay verified the targeting relationship between circGPX1 and miR-150.The expression of miR-150 in TPC-1 cells of 2 groups was detected by qRT-PCR.The expressions of Wnt/β-catenin signaling pathway proteinsβ-catenin,c-myc,p-GSK3β and cyclin D in TPC-1 cells of 2 groups were detected by using Western blot.Results In the GeneCards database,the transcriptional level of circGPX1 is highly expressed in thyroid cancer tissues compared with paracancerous tissues,and the difference was statistically significant(P<0.01);the relative expression level of circGPX1 was increased with the higher clinical stage of thyroid cancer patients,and the difference was statistically significant(P<0.01).Compared with Nthy-ori3-1 cells,the transcriptional l

关 键 词：甲状腺肿瘤 RNA 环状 细胞增殖 肿瘤侵润 circGPX1 miRNA-150 

分 类 号：R736.1[医药卫生—肿瘤]