环状RNA锌指蛋白652靶向微小RNA-1278调控叉头盒P1蛋白表达对食管鳞状细胞癌细胞存活的影响  

Effects of circularRNA zinc finger protein 652 targeting microRNA-1278 to regulate forkhead box P1 expression on cell survival of esophageal squamous cell carcinoma

作　　者：周昆[1] 闫焱[2] 李冰[1] 赵志超 Zhou Kun;Yan Yan;Li Bing;Zhao Zhichao(Department of Thoracic Surgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Department of Oncology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院胸外科,450052 [2]郑州大学第一附属医院肿瘤科,450052

出　　处：《中华实验外科杂志》2022年第7期1314-1320,共7页Chinese Journal of Experimental Surgery

基　　金：河南省高等学校重点科研项目(20A320065);河南省医学科技攻关计划项目(201702010)。

摘　　要：目的探讨环状RNA锌指蛋白652(circZNF652)/微小RNA(miR)-1278/叉头盒P1蛋白(FOXP1)轴对食管鳞状细胞癌(ESCC)细胞活性、增殖、凋亡和侵袭能力的影响。方法实时荧光定量聚合酶链反应(qPCR)检测circZNF652、miR-1278以及FOXP1 mRNA表达;细胞计数试剂盒(CCK-8)实验检测细胞活性;EdU实验检测细胞增殖;流式细胞仪检测细胞凋亡;Transwell实验检测细胞侵袭;蛋白质印迹法(Western blot)实验检测FOXP1蛋白表达;双荧光素酶报告基因实验和RNA免疫共沉淀(RIP)实验验证circZNF652与miR-1278之间的靶向关系,以及miR-1278与FOXP1之间的靶向关系。以t检验或单因素方差分析进行统计分析。结果ESCC细胞系KYSE-510和TE-10中circZNF652、FOXP1 mRNA和蛋白表达高于正常食管上皮细胞HEEC(3.37±0.24比1.97±0.06、4.38±0.29比2.88±0.12、2.95±0.25比1.48±0.12),差异有统计学意义(F=209.4、267.3、75.5,P<0.05),而KYSE-510和TE-10中miR-1278表达低于HEEC(0.47±0.06比0.71±0.03),差异有统计学意义(F=152.5,P<0.05)。circZNF652小干扰RNA(si-circZNF652)组的细胞活性、增殖和侵袭能力低于阴性对照小干扰RNA(si-NC)组,差异有统计学意义(0.44±0.04比0.71±0.03、0.50±0.04比1.05±0.05、0.46±0.05比1.01±0.07),差异有统计学意义(F=1.5、1.3、2.1,P<0.05);而si-circZNF652组的细胞凋亡率高于si-NC组[(30.2±0.28)%比(5.20±1.27)%],差异有统计学意义(F=2.3,P<0.05)。miR-1278拮抗剂(抗miR-1278)组的细胞活性、增殖和侵袭能力高于阴性对照拮抗剂(抗NC)组(1.02±0.03比0.71±0.03、1.38±0.04比1.04±0.05、1.51±0.06比0.99±0.04),差异有统计学意义(F=484.1、306.1、190.6,P<0.05),而si-circZNF652+抗miR-1278组的细胞活性、增殖和侵袭能力高于si-circZNF652+抗NC组(0.57±0.02比0.35±0.02、0.89±0.04比0.49±0.02、0.88±0.04比0.52±0.06),差异有统计学意义(F=484.1、306.1、190.6,P<0.05)。miR-1278 mimic+FOXP1过表达(OE-FOXP1)组的细胞活性、增殖和侵袭能力高于miR-1278 mimic+FOXPObjective To investigate the effects of circularRNA zinc finger protein 652(circZNF652)/microrNA(miR)-1278/forkhead box P1(FOXP1)axis on the cell viability,proliferation,apoptosis and invasion of esophageal squamous cell carcinoma(ESCC)cells.Methods The expression of circZNF652,miR-1278 and FOXP1 mRNA was detected by quantitative real-time polymerase chain reaction(qPCR).Cell viability was determined by cell counting kit-8(CCK-8).Cell proliferation was detected by EdU assay.Cell apoptosis was measured by flow cytometry.Cell invasion was analyzed by Transwell assay.FOXP1 protein expression was detected by Western blotting.The dual luciferase reporter gene assay and RNA immunoprecipitation(RIP)assay were used to verify the targeting relationship between circZNF652 and miR-1278,as well as the targeting relationship between miR-1278 and FOXP1.T test or one-way ANOVA was used for statistical analysis.Methods The expression of circZNF652,and FOXP1 mRNA and protein in ESCC cell lines KYSE-510 and TE-10 were higher than those in normal esophageal epithelial cells HEEC(3.37±0.24 vs.1.97±0.06,4.38±0.29 vs.2.88±0.12,2.95±0.25 vs.1.48±0.12),the differences were statistically significant(F=209.4,267.3,75.5,P<0.05),and miR-1278 expression in KYSE-510 and TE-10 was lower than that in HEEC(0.47±0.06 vs.0.71±0.03),and the difference was statistically significant(F=152.5,P<0.05).The cell viability,proliferation and invasion ability of circZNF652 small interference RNA(si-circZNF652)group were lower than that of negative control-small interference RNA(si-NC)group,and the differences were statistically significant(0.44±0.04 vs.0.71±0.03,0.50±0.04 vs.1.05±0.05,0.46±0.05 vs.1.01±0.07),the differences were statistically significant(F=1.5,1.3,2.1,P<0.05);The apoptosis rate of si-circZNF652 group was higher than that of Si-NC group[(30.2±0.28)%vs.(5.20±1.27)%],and the difference was statistically significant(F=2.3,P<0.05).The cell activity,proliferation and invasions of mir-1278 antagomir(anti-miR-1278)group were higher t

关 键 词：食管癌 环状ZNF652 微小RNA 细胞存活 

分 类 号：R735.1[医药卫生—肿瘤]