miR-450b-5p调控IL-1β诱导人牙龈上皮细胞粘附和迁移  

miR-450b-5p Regulates Adhesion and Migration of Human Gingival Epithelial Cells Induced by IL-1β

作　　者：禹翠翠 叶潇元 董志恒[1] 王爱芹[2] 高艳[1] 高玉光[1] YU Cuicui;YE Xiaoyuan;DONG Zhiheng;WANG Aiqin;GAO Yan;GAO Yuguang(Department of Pediatric Dentistry,Hospital of Binzhou Medical University,Binzhou 256600,China;Department of Oral Medicine,Hospital of Binzhou Medical University,Binzhou 256600,China)

机构地区：[1]滨州医学院附属医院儿童口腔科,山东滨州256600 [2]滨州医学院附属医院口腔内科,山东滨州256600

出　　处：《口腔医学研究》2022年第6期572-577,共6页Journal of Oral Science Research

基　　金：国家自然科学基金(编号:81870738)。

摘　　要：目的:本研究旨在明确miR-450b-5p对炎症因子IL-1β调控人牙龈上皮细胞(HGEs)粘附与迁移的作用。方法:体外培养HGEs,在对数生长期饥饿培养12 h,5 ng/mL IL-1β刺激3 h,RNAiso Plus提取总RNA,进行small RNA测序以及实时荧光定量-聚合酶链反应(qRT-PCR)分析;细胞划痕实验观察IL-1β调控HGEs迁移;转染miR-450b-5p mimic、inhibitor和NC慢病毒质粒,qRT-PCR检测IL-1β调控HGEs中miR-450b-5p、Integrinβ6(ITGβ6)、Lamininγ2(LAMC2)和基质金属蛋白酶-9(MMP-9)的表达。结果:测序结果和qRT-PCR示IL-1β诱导miR-450b-5p表达降低;IL-1β诱导ITGβ6、LAMC2和MMP-9表达升高;过表达miR-450b-5p抑制ITGβ6、LAMC2和MMP-9基因表达,部分挽救IL-1β诱导的ITGβ6、LAMC2和MMP-9表达升高,敲低miR-450b-5p则出现相反结果;细胞划痕实验示过表达miR-450b-5p抑制IL-1β诱导的HGEs迁移。结论:miR-450b-5p抑制IL-1β诱导HGEs粘附与迁移,提示miR-450b-5p对维持HGEs稳定、抑制细胞迁移以及保护牙龈上皮屏障有重要作用。Objective:To investigate the effect of miR-450b-5p on the adhesion and migration of human gingival epithelial cells(HGEs)regulated by inflammatory factor IL-1β.Methods:HGEs were cultured in vitro and starved for 12 hours in logarithmic growth period,stimulated by 5 ng/mL IL-1βfor 3 hours.Total RNA was extracted by RNAiso Plus and analyzed by small RNA sequencing and qRT-PCR.The migration of HGEs induced by IL-1βwas observed by cell scratch assay.After transfected with miR-450b-5p mimic,inhibitor,and NC lentivirus plasmid,the expressions of miR-450b-5p,Integrinβ6(ITGβ6),Lamininγ2(LAMC2),and matrix metalloproteinase-9(MMP-9)in HGEs regulated by IL-1βwere detected by qRT-PCR.Results:Sequencing results and qRT-PCR showed that IL-1βdecreased the expression of miR-450b-5p.The expression of ITGβ6,LAMC2,and MMP-9 were increased after IL-1βinduction.The expressions of ITGβ6,LAMC2,and MMP-9 genes were inhibited and the increased effect induced by IL-1βwas partially reversed by overexpression of miR-450b-5p.However,the opposite result was obtained by knockdown of miR-450b-5p.Cell scratch assay showed that migration of HGEs induced by IL-1βwas inhibited by overexpression of miR-450b-5p.Conclusion:MiR-450b-5p inhibits HGEs adhesion and migration induced by IL-1β,suggesting that miR-450b-5p plays an important role in maintaining HGEs stability,inhibiting cell migration and protecting the gingival epithelial barrier.

关 键 词：白细胞介素-1Β 微小RNA-450b-5p 层粘连蛋白γ2 整合素β6 基质金属蛋白酶-9 

分 类 号：R73[医药卫生—肿瘤]