circIRAK3靶向miR-942-5p/LITAF轴调控滋养层细胞的增殖、迁移、侵袭和EMT  

circIRAK3Targets miR-942-5p/LITAF Axis to Regulate Proliferation,Migration,Invasion and EMT of Trophoblastic Cells

作　　者：李蕊[1] 林靓[1] 胡绪洋 翁晓英 王军军 Li Rui;Lin Liang;Hu Xuyang(Department of Obstetrics and Gynecology,Shengli Clinical Medical College of Fujian Medical University,Fuzhou 350500,China)

机构地区：[1]福建医科大学省立临床医学院妇产科,福州350500 [2]福建医科大学省立临床医学院检验科,福州350500

出　　处：《华中科技大学学报（医学版）》2023年第5期603-609,共7页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：福建省自然科学基金资助项目(No.2020J011066)。

摘　　要：目的探讨环状RNA circIRAK3对滋养层细胞生物学行为的影响及其潜在分子调控机制。方法采集19例正常足月妊娠孕妇和25例早发型子痫前期(PE)孕妇的血液和胎盘组织样本,实时荧光定量PCR(qRT-PCR)分析circIRAK3的表达量,受试者工作特征曲线(ROC)分析其对PE的诊断价值。体外培养人滋养层细胞HTR-8/SVneo,通过CCK-8实验、划痕实验和Transwell实验评估细胞的增殖、迁移和侵袭能力;Western blot实验检测上皮间充质转化(EMT)相关蛋白表达。采用生物信息学方法和双荧光素酶报告基因实验分析并验证circIRAK3的调控机制。结果circIRAK3在PE患者血液和胎盘组织样本中高表达,且血液中circIRAK3的表达对PE具有一定诊断价值(AUC=0.882)。细胞功能研究显示,过表达circIRAK3能够抑制HTR-8/SVneo细胞的增殖、迁移和侵袭。调控机制分析显示,miR-942-5p是circIRAK3的靶miRNA,而脂多糖诱导肿瘤坏死因子α(LITAF)是miR-942-5p的靶基因,circIRAK3可作为miR-942-5p的分子海绵上调LITAF的表达;过表达miR-942-5p或抑制LITAF表达均能减弱circIRAK3对HTR-8/SVneo细胞增殖、迁移和侵袭的抑制作用。结论circIRAK3通过靶向miR-942-5p上调LITAF的表达,从而抑制HTR-8/SVneo细胞的增殖、迁移和侵袭,提示circIRAK3可能参与PE的发病。Objective To investigate the effect of circIRAK3on the biological behavior of trophoblast cells,and its potential molecular regulation mechanism.Methods Blood and placental tissue samples were collected from 19healthy pregnant women and 25pregnant women with preeclampsia(PE).The expression of circIRAK3was analyzed by qRT-PCR,and the diagnostic value of circIRAK3for PE was analyzed by receiver operating characteristic curve(ROC).Human trophoblast cells(HTR-8/SVneo)were cultured in vitro and evaluated for proliferation,migration and invasion by CCK-8assay,scratch assay and Transwell assay.The expression of epithelial-mesenchymal transformation(EMT)related proteins was detected by Western blotting.Bioinformatics analysis and dual luciferase assay were used to analyze and verify the regulatory mechanism of circIRAK3.Results circIRAK3was highly expressed in blood and placental tissue samples of PE patients,and the expression of circIRAK3in blood had certain diagnostic value for PE(AUC=0.882).Functional studies showed that overexpression of circIRAK3could inhibit the proliferation,migration and invasion of HTR-8/SVneo cells.Analysis of regulatory mechanism showed that miR-942-5p was the target miRNA of circIRAK3,while LITAF was the target gene of miR-942-5p,and circIRAK3could act as a sponge for miR-942-5p to up-regulate the expression of LITAF.Overexpression of miR-942-5p or inhibition of LITAF weakened the inhibitory effect of circIRAK3overexpression on proliferation,migration and invasion of HTR-8/SVneo cells.Conclusion circIRAK3upregulated LITAF expression by targeting miR-942-5p,thus inhibiting the proliferation,migration and invasion of HTR-8/SVneo cells,suggesting that circIRAK3may be involved in the pathogenesis of PE.

关 键 词：子痫前期 circIRAK3 人滋养层细胞 miR-942-5p LITAF 

分 类 号：R714.244[医药卫生—妇产科学]