miR-216靶向调控HMGA2对宫颈癌细胞侵袭和迁移的影响  

Effect of miR-216 targeting HMGA2 on invasion and migration of cervical cancer cells

作　　者：曾康康 郑蓉[1] 刘永珍[1] 贺敏[1] ZENG Kangkang;ZHENG Rong;LIU Yongzhen;HE Min(Shiyan Taihe Hospital of Hubei Province,Shiyan 442700,China)

机构地区：[1]十堰市太和医院

出　　处：《山东医药》2019年第31期14-18,共5页Shandong Medical Journal

摘　　要：目的探讨微小RNA-216(miR-216)靶向调控高迁移率族蛋白A2(HMGA2)对宫颈癌细胞侵袭和迁移的影响。方法采用RT-qPCR法检测人宫颈癌细胞SiHa、HeLa、CaSki和人正常宫颈上皮细胞H8中miR-216表达,选取miR-216表达最低的宫颈癌细胞进行后续实验。将miR-216表达最低的宫颈癌细胞随机分为观察组与对照组,分别转染miR-216 mimics、miR-216 NC。转染24 h,收集细胞,采用RT-qPCR法检测转染效率,采用Transwell迁移实验和Boyden侵袭实验检测细胞迁移和侵袭能力,采用Western blotting法检测MMP-2、MMP-9蛋白表达。采用TargetScan软件预测HMGA23′UTR与miR-216的结合位点。取上述miR-216表达最低的宫颈癌细胞,随机分为miR-216 mimics+HMGA2 WT组、miR-216 mimics+HMGA2 MUT组、miR-216 NC+HMGA2 WT组、miR-216 NC+HMGA2 MUT组,miR-216 mimics+HMGA2 WT组转染miR-216 mimics和HMGA2 WT,miR-216 mimics+HMGA2 MUT组转染miR-216 mimics和HMGA2 MUT,miR-216 NC+HMGA2 WT组转染miR-216 NC和HMGA2 WT,miR-216 NC+HMGA2 MUT组转染miR-216 NC和HMGA2 MUT。转染24 h,收集细胞,采用双荧光素酶报告基因实验检测荧光素酶活性。结果HeLa、SiHa、CaSki细胞miR-216相对表达量均明显低于H8细胞(P均<0.05)。在宫颈癌细胞中以HeLa细胞miR-216相对表达量最低,故选择HeLa细胞进行后续实验。观察组miR-216相对表达量明显高于对照组(P<0.05)。Transwell迁移实验和Boyden侵袭实验时,观察组穿膜细胞数均低于对照组(P均<0.05)。观察组MMP-2、MMP-9蛋白相对表达量均低于对照组(P均<0.05)。TargetScan软件预测结果显示,HMGA2是miR-216的靶基因。双荧光素酶报告基因实验显示,miR-216 mimics+HMGA2 WT组荧光素酶活性低于miR-216 NC+HMGA2 WT组(P<0.05),而miR-216 mimics+HMGA2 MUT组与miR-216 NC+HMGA2 MUT组荧光素酶活性比较P>0.05。结论miR-216能够通过靶向调控HMGA2抑制宫颈癌细胞侵袭和迁移。Objective To investigate the effect of miR-216 targeting high mobility group box A2(HMGA2)on the invasion and migration of cervical cancer cells.Methods The expression of miR-216 in the human cervical cancer cells SiHa,HeLa,CaSki and human normal cervical epithelial cells H8 were detected by qRT-PCR.The cervical cancer cells with the lowest expression of miR-216 were selected for subsequent experiments.Cervical cancer cells with the lowest expression of miR-216 were randomly divided into the observation group and control group,and transfected with miR-216 mimics and miR-216 NC,respectively.The cells were harvested at 24 h,and the transfection efficiency was detected by qRT-PCR.Transwell assay and Boyden assay were used to detect cell migration and invasion.The expression of matrix metalloproteinase 2(MMP-2)and MMP-9 protein was detected by Western blotting.The binding site of HMGA23′UTR to miR-216 was predicted using TargetScan software.Human cervical cancer cells with the lowest expression of miR-216 were randomly divided into the miR-216 mimics+HMGA2 WT group,miR-216 mimics+HMGA2 MUT group,miR-216 NC+HMGA2 WT group,and miR-216 NC+HMGA2 MUT group;cells in the miR-216 mimics+HMGA2 WT group were transfected with miR-216 mimics and HMGA2 WT,miR-216 mimics+HMGA2 MUT group with miR-216 mimics and HMGA2 MUT,miR-216 NC+HMGA2 WT group with miR-216 NC and HMGA2 WT,and miR-216 NC+HMGA2 MUT group with miR-216 NC and HMGA2 MUT;they were all transfected for 24 h,and the luciferase activity was detected by a dual luciferase reporter assay.Results The relative expression of miR-216 in HeLa,SiHa and CaSki cells was significantly lower than that in H8 cells(P<0.05).Among them,the relative expression of miR-216 in HeLa cells was the lowest in cervical cancer cells,so HeLa cells were selected for the subsequent experiments.The relative expression of miR-216 in the observation group was significantly higher than that in the control group(P<0.05).In the Transwell assay and the Boyden assay,the number of trans-membrane cells in the

关 键 词：宫颈癌 微小RNA-216 高迁移率族蛋白A2 细胞侵袭 细胞迁移 

分 类 号：R737.33[医药卫生—肿瘤]