长链非编码RNA-GAS5竞争性结合miR-10调控SiHa细胞的增殖、迁移及侵袭能力的影响  

Effects of long non-coding RNA-GAS5 competitively binding miR-10 on the regulation of proliferation,migration and invasion ability of SiHa cells

作　　者：王阳阳 刘耘 杨红梅 董仙萍 刘桂艳 WANG Yangyang;LIU Yun;YANG Hongmei;DONG Xianping;LIU Guiyan(Department of Gynaecology,Tangshan Central Hospital,Tangshan,Hebei 067000,China;Department of Gynaecology,Kailuan General Hospital,Tangshan,Hebei 067000,China;Department of Gynaecology,Tangshan Women's and Children's Hospital,Tangshan,Hebei 067000,China)

机构地区：[1]河北省唐山中心医院妇科,河北唐山067000 [2]河北省开滦总医院妇科,河北唐山067000 [3]河北省唐山市妇幼保健院妇科,河北唐山067000

出　　处：《检验医学与临床》2023年第23期3453-3457,共5页Laboratory Medicine and Clinic

基　　金：河北省医学科学研究重点课题资助项目(20210127)。

摘　　要：目的探讨长链非编码RNA(lncRNA)-生长停滞特异性转录5(GAS5)竞争性结合微小RNA-10(miR-10)调控人子宫颈鳞癌(SiHa)细胞的增殖、迁移及侵袭能力的影响。方法选取2021年1月至2022年1月在河北省唐山中心医院接受手术治疗的48例宫颈癌手术患者作为研究对象,提取所有研究对象的肿瘤样本和癌旁正常组织样本中的SiHa细胞,按照转染方式的不同将培养的SiHa细胞分为对照组、GAS5组、miR-10组、血小板反应蛋白(THBS)2组、GAS5+miR-10组和GAS5+miR-10+THBS2组。检测lncRNA-GAS5、miR-10和THBS2在其肿瘤样本和癌旁正常组织样本中的表达水平;采用Starbase、TargetScan数据库预测lncRNA-GAS5和miR-10及miR-10和THBS2的结合位点;检测转染lncRNA-GAS5和THBS2后SiHa细胞的吸光度(A)及侵袭细胞数。结果宫颈癌组织的miR-10表达水平高于癌旁正常组织,而宫颈癌组织的lncRNA-GAS5、THBS2表达水平低于癌旁正常组织,差异均有统计学意义(P<0.05)。将lncRNA-GAS5过表达质粒转染进SiHa细胞后,lncRNA-GAS5组miR-10表达水平较对照组明显升高(P<0.05)。转染miR-10后,miR-10组SiHa细胞中的miR-10表达水平比转染前高(P<0.05)。lncRNA-GAS5的3′-UTR有miR-10的结合位点,miR-10在THBS2的3′-UTR上具有潜在的结合位点。相比对照组,miR-10组PMIR-GAS5-WT及PMIR-THBS2-WT野生型质粒荧光素酶活性明显降低(P<0.05)。相比对照组,GAS5组和THBS2组SiHa细胞的A及侵袭细胞数降低,而miR-10组SiHa细胞的A及侵袭细胞数升高,差异均有统计学意义(P<0.05);相比miR-10组,GAS5+miR-10组SiHa细胞的A和细胞迁移细胞数降低,差异均有统计学意义(P<0.05);相比GAS5+miR-10组,GAS5+miR-10+THBS2组SiHa细胞的A和细胞迁移细胞数降低,差异均有统计学意义(P<0.05)。结论lncRNA-GAS5能通过抑制miR-10上调THBS2的表达水平从而抑制宫颈癌细胞的生物学行为。Objective To investigate the effects of long non-coding RNA(lncRNA)-growth arrest-specific 5(GAS5)competitively binding microRNA-10(miR-10)on the regulation of proliferation,migration and invasion ability of human cervical squamous cell carcinoma(SiHa)cells.Methods A total of 48 patients with cervical cancer who underwent surgery in Tangshan Central Hospital of Hebei Province from January 2021 to January 2022 were selected as the research objects.SiHa cells were extracted from the tumor samples and adjacent normal tissue samples of all the research objects.According to different transfection methods,SiHa cells were divided into control group,GAS5 group,miR-10 group,thrombospondin(THBS)2 group,GAS5+miR-10 group and GAS5+miR-10+THBS2 group.The expression levels of lncRNA-GAS5,miR-10 and THBS2 in tumor samples and adjacent normal tissue samples were detected.The Starbase and TargetScan databases were used to predict the binding sites of lncRNA-GAS5,miR-10 and miR-10,THBS2.The optical density(A)of SiHa cells transfected with lncRNA-GAS5 and THBS2 and the number of invasive cells were detected.Results The expression level of miR-10 in cervical cancer tissues was higher than that in adjacent normal tissues,while the expression levels of lncRNA-GAS5 and THBS2 in cervical cancer tissues were lower than those in adjacent normal tissues,and the differences were statistically significant(P<0.05).After the lncRNA-GAS5 overexpression plasmid was transfected into SiHa cells,the miR-10 expression level of lncRNA-GAS5 group was significantly higher than that of the control group(P<0.05).After transfection with miR-10,the miR-10 expression level of SiHa cells in the miR-10 group was higher than that before transfection(P<0.05).lncRNA-GAS5 had a miR-10 binding site in the 3'-UTR,and miR-10 had a potential binding site in the 3'-UTR of THBS2.Compared with the control group,the luciferase activities of PMIR-GAS5-WT and PMIR-THBS2-WT wild-type plasmids in the miR-10 group were significantly decreased(P<0.05).Compared with the control

关 键 词：宫颈癌 生长停滞特异性转录5 微小RNA-10 血小板反应蛋白2 

分 类 号：R737.33[医药卫生—肿瘤]