微小RNA-558靶向调控叉头框转录因子C1促进卵巢癌SKOV3细胞的增殖和侵袭力  

MicroRNA-558 promotes the proliferation and invasion of SKOV3 cells by targeting FOXC1

作　　者：李静[1] 王金铭 任琛琛[1] 杨立[1] 刘泇希 白杨[1] LI Jing;WANG Jinming;REN Chenchen;YANG Li;LIU Jiaxi;BAI Yang(Department of Obstetrics and Gynecology,the Third Affiliated Hospital of Zhengzhou University,Zhengzhou,Henan 450000;Institute of Medical Genetics,Henan Provincial People′s Hospital,Zhengzhou,Henan 450000)

机构地区：[1]郑州大学第三附属医院妇产科,河南郑州450000 [2]河南省人民医院医学遗传研究所,河南郑州450000

出　　处：《安徽医药》2020年第10期1943-1947,共5页Anhui Medical and Pharmaceutical Journal

摘　　要：目的探讨miR-558通过对叉头框转录因子C1(Forkhead box protein C1,FOXC1)的调控从而对人卵巢癌腺癌细胞(SKOV3)增殖和侵袭能力的影响。方法选择2014年1月至2017年12月郑州大学第三附属医院妇产科收治的上皮性卵巢癌病人25例、交界性卵巢上皮性肿瘤病人25例和良性卵巢上皮性肿瘤病人25例,三组病例均行手术治疗,取其部分经手术切除的卵巢组织标本,使用实时荧光定量PCR技术来分析微小RNA-558(miR-558)的表达水平。把卵巢癌SKOV3细胞分成三组,分别为miR-558模拟物组、抑制物组和阴性对照组。通过靶基因预测网站来预测miR-558的靶基因(FOXC1基因),通过荧光素酶报告基因实验来验证miR-558对FOXC1基因表达的调控作用。通过实时荧光定量-PCR技术和蛋白印迹法来分析转染后各组细胞的miR-558和FOXC1的表达水平。通过细胞增殖实验(Cell Counting Kit-8,CCK-8法)检测三组细胞的增殖率。通过基质胶侵袭实验检测三组细胞的侵袭能力。结果实时荧光定量-PCR结果显示,在上皮性卵巢癌、交界性卵巢上皮性肿瘤和良性卵巢上皮性肿瘤病人的卵巢组织中,miR-558的相对表达水平分别为(3.43±0.42)、(2.47±0.35)、(1.37±0.31);在miR-558模拟物组、抑制物组和阴性对照组中,SKOV3细胞miR-558的表达水平分别为(2.37±0.17)、(0.64±0.17)、(1.14±0.11)。在miR-558模拟物组、抑制物组和阴性对照组中,SKOV3细胞的FOXC1基因转录出的信使RNA(mRNA)表达水平分别为(0.51±0.10)、(2.27±0.12)、(0.99±0.11)。荧光素酶报告基因实验结果显示,SKOV3细胞被含miR-558的质粒以及含FOXC1基因的重组质粒共同转染后,的荧光素酶活性下降了49.50%(P<0.05)。蛋白印迹结果显示,上述三组SKOV3细胞中,FOXC1蛋白的表达水平分别为(0.83±0.07)、(2.17±0.15)、(1.47±0.21)。CCK-8检测结果显示,不同时间miR-558模拟物组SKOV3细胞的增殖率明显高于阴性对照组(P<0.05),不同时间miR-558�Objective To clarify the role of miR-558 on the proliferation and invasion of SKOV3 cell and the relationship withFOXC1.MethodsWe recruited 25 patients with epithelial ovarian cancer,25 patients with borderline epithelial ovarian tumorsand 25 patients with benign epithelial ovarian tumors in Department of Obstetrics and Gynecology of the Third Affiliated Hospitalof Zhengzhou University from January 2014 to December 2017.The three groups underwent surgical treatment,and some ovarian tis-sue of patients were taken.Quantitative real-time polymerase chain reaction(q RT-PCR)was used to measure the expression of mi R-558 in ovarian tissues.SKOV3 cells were assigned to mi R-558 mimics group,inhibitor group,negative control,respectively.Bioinfor-matics was used for predicting target genes for mi RNA-558-5 p(FOXC1)and dual-luciferase reporting system for verifying the tar-get gene.Mi R-558 expression was evaluated by q RT-PCR.FOXC1 expression was evaluated by western blot and q RT-PCR.Cell pro-liferation was determined by CCK-8 assay.Invasive activities were assessed by cell Transwell invasion assay.ResultsThe relativeexpression of mi R-558 measured by q RT-PCR in the ovarian tissues of patients with epithelial ovarian cancer,borderline ovarianepithelial tumor and benign ovarian epithelial tumor were(3.43±0.42),(2.47±0.35),(1.37±0.31),respectively.The expression of mi R-558 in SKOV3 cell line in the mi R-558 mimics group,inhibitor group and negative control group were(2.37±0.17),(0.64±0.17),(1.14±0.11),respectively.The expression of FOXC1 m RNA in SKOV3 cell line in the mi R-558 mimics group,inhibitorgroup and negative control group were(0.51±0.10),(2.27±0.12),(0.99±0.11),respectively.Luciferase reporter assay resultsshowed that the luciferase activity of SKOV3 cells transfected with mi R-558 containing plasmid and FOXC1 containing recombi-nant plasmid decreased by 49.50%(P<0.05).The expression of FOXC1 protein in SKOV3 cell line in the mi R-558 mimics group,inhibitor group and negative control group were(0.83±0

关 键 词：卵巢肿瘤 叉头转录因子类 微小RNA 增殖 实时聚合酶链反应 荧光免疫测定 实时荧光定量PCR 侵袭 

分 类 号：R737.31[医药卫生—肿瘤]