lncRNA TFAP2A-AS1/miR-9-5p通过ERK通路对子宫内膜癌细胞增殖、侵袭及迁移的影响  

The effects of lncRNA TFAP2A-AS1/miR-9-5p on proliferation,invasion and migration of endometrial carcinoma cells through ERK pathway

作　　者：赵艳宏 赵达 傅琳 苏丹 郭博 ZHAO Yanhong;ZHAO Da;FU Lin;SU Dan;GUO Bo(Hainan Women and Children Medical Center,Hainan Haikou 570100,China)

机构地区：[1]海南省妇女儿童医学中心,海南海口570100

出　　处：《现代肿瘤医学》2024年第1期18-24,共7页Journal of Modern Oncology

基　　金：海南省临床医学中心建设项目资助(编号:琼卫医函[2021]75号)。

摘　　要：目的:探讨长链非编码RNA TFAP2A-AS1对子宫内膜癌细胞增殖、侵袭和迁移的影响及机制。方法:选择50例子宫内膜癌组织和对应的癌旁组织。选取子宫内膜癌细胞株(RL95-2、HEC-1A、HHUA、HEC-1B及Ishikawa),子宫内膜上皮细胞hEEC。子宫内膜癌细胞株Ishikawa转染si-TFAP2A-AS1(si-TFAP2A-AS1组)、si-NC(si-NC组)、miR-9-5p mimics(miR-9-5p组)及miR-NC(miR-NC组);RL95-2细胞转染OE-TFAP2A-AS1(TFAP2A-AS1组)、Vector(Vector组)、sh-miR-9-5p(sh-miR-9-5p组)及sh-NC(sh-NC组)。CCK-8检测细胞增殖能力,Transwell检测细胞侵袭、迁移能力,双荧光素酶实验、pull down实验分析TFAP2A-AS1与miR-9-5p的靶向关系;miR-9-5p与ERK的靶向关系,实时荧光定量PCR(RT-qPCR)检测子宫内膜癌组织、癌旁组织、子宫内膜癌细胞及hEEC细胞TFAP2AAS1、miR-9-5p表达水平,Western blot检测细胞外调节蛋白激酶(ERK)、磷酸化ERK(p-ERK)水平。结果:子宫内膜癌组织中TFAP2A-AS1表达水平低于癌旁组织,miR-9-5p表达水平高于癌旁组织(P<0.05)。子宫内膜癌组织TFAP2A-AS1、miR-9-5p表达水平与分化程度、肿瘤直径、TNM分期及淋巴结转移有关。子宫内膜癌组织中TFAP2A-AS1、miR-9-5p表达水平呈负相关(r=-0.782,P=0.002)。与hEEC细胞比较,RL95-2、HEC-1A、HHUA、HEC-1B及Ishikawa细胞TFAP2A-AS1表达水平降低,miR-9-5p表达水平升高(P<0.05)。与si-NC组比较,si-TFAP2A-AS1组细胞48 h、72 h OD值,侵袭、迁移细胞数均升高(P<0.05);与Vector组比较,TFAP2A-AS1组细胞48 h、72 h OD值,侵袭、迁移细胞数均降低(P<0.05)。与miR-NC组比较,miR-9-5p组细胞48 h、72 h OD值,侵袭、迁移细胞数均升高(P<0.05);与sh-NC组比较,sh-miR-9-5p组细胞48 h、72 h OD值,侵袭、迁移细胞数均降低(P<0.05)。TFAP2A-AS1负性调控miR-9-5p、p-ERK表达水平,miR-9-5p可靶向调控ERK蛋白。结论:过表达TFAP2A-AS1通过靶向下调miR-9-5p,抑制ERK通路,最终抑制子宫内膜癌细胞恶性生物学行为。Objective:To investigate the effect and mechanism of lncRNA TFAP2A-AS1 on the proliferation,invasion and migration of endometrial cancer cells.Methods:50 cases of endometrial carcinoma tissues and corresponding adjacent tissues were selected.Endometrial carcinoma cell lines(RL95-2,HEC-1A,HHUA,HEC-1B and Ishikawa) and endometrial epithelial cells hEEC were selected.Ishikawa transfected si-TFAP2A-AS1(si-TFAP2A-AS1 group),si-NC(si-NC group),miR-9-5p mimics(miR-9-5p group) and miR-NC(miR-NC group).RL95-2 cells were transfected with OE-TFAP2A-AS1(TFAP2A-AS1 group),Vector(Vector group),sh-miR-9-5p(sh-miR-9-5p group) and sh-NC(sh-NC group).CCK-8 was used to detect cell proliferation.Transwell was used to detect cell invasion and migration.Double luciferase and pull down assays were used to analyze the targeting relationship between TFAP2A-AS1 and miR-9-5p and the targeting relationship between miR-9-5p and ERK.The expression levels of TFAP2A-AS1 and miR-9-5p in endometrial carcinoma tissues,adjacent tissues,endometrial carcinoma cells,and hEEC cells were detected by real-time fluorescent quantitative PCR(RT-qPCR).The levels of extracellular regulated protein kinase(ERK) and phosphorylated ERK(p-ERK)were detected by Western blot.Results:The expression level of TFAP2A-AS1 in endometrial carcinoma was lower than that in adjacent tissues,while the expression level of miR-9-5p was higher than that in adjacent tissues(P<0.05).The expression level of TFAP2A-AS1 and miR-9-5p in endometrial carcinoma is related to the degree of differentiation,tumor diameter,TNM stage and lymph node metastasis.The expression levels of TFAP2A-AS1 and miR-9-5p in endometrial carcinoma were negatively correlated(r=-0.782,P= 0.002).Compared with hEEC cells,the expression level of TFAP2A-AS1 in RL95-2,HEC-1A,HHUA,HEC-1B and Ishikawa cells decreased,and the expression level of miR-9-5p increased(P< 0.05).Compared with si-NC group,the OD value of cells in si-TFAP2A-AS1 group at 48 h and 72 h,and the number of invasive and migratory cells increased(P<0.

关 键 词：子宫内膜癌 长链非编码RNA TFAP2AAS1 miR-9-5p ERK通路 

分 类 号：R737.33[医药卫生—肿瘤]