miR-450b-5p调控IL-1β诱导人牙龈上皮细胞粘附和迁移

doi:10.13701/j.cnki.kqyxyj.2022.06.016

口腔医学研究 ›› 2022, Vol. 38 ›› Issue (6): 572-577.DOI: 10.13701/j.cnki.kqyxyj.2022.06.016

miR-450b-5p调控IL-1β诱导人牙龈上皮细胞粘附和迁移

禹翠翠¹, 叶潇元¹, 董志恒¹, 王爱芹^2*, 高艳^1*, 高玉光¹

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

收稿日期:2022-01-11 出版日期:2022-06-28 发布日期:2022-06-23
通讯作者: * 王爱芹,E-mail:bywaq@126.com;高艳,E-mail:gaoyanbinyi@sina.com
作者简介:禹翠翠(1985~),女,山东阳信人,主治医师,硕士在读,研究方向:儿童口腔医学。
基金资助:
国家自然科学基金(编号:81870738)

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

YU Cuicui¹, YE Xiaoyuan¹, DONG Zhiheng¹, WANG Aiqin^2*, GAO Yan^1*, GAO Yuguang¹

1. Department of Pediatric Dentistry, Hospital of Binzhou Medical University, Binzhou 256600, China;
2. Department of Oral Medicine, Hospital of Binzhou Medical University, Binzhou 256600, China

Received:2022-01-11 Online:2022-06-28 Published:2022-06-23

摘要/Abstract

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。

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

Abstract: 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-1β for 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-1β was 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-1β were detected by qRT-PCR. Results: Sequencing results and qRT-PCR showed that IL-1β decreased the expression of miR-450b-5p. The expression of ITGβ6, LAMC2, and MMP-9 were increased after IL-1β induction. The expressions of ITGβ6, LAMC2, and MMP-9 genes were inhibited and the increased effect induced by IL-1β was 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-1β was 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.

Key words: IL-1β, miR-450b-5p, lamininγ2, ITGβ6, MMP-9

禹翠翠, 叶潇元, 董志恒, 王爱芹, 高艳, 高玉光. miR-450b-5p调控IL-1β诱导人牙龈上皮细胞粘附和迁移[J]. 口腔医学研究, 2022, 38(6): 572-577.

YU Cuicui, YE Xiaoyuan, DONG Zhiheng, WANG Aiqin, GAO Yan, GAO Yuguang. miR-450b-5p Regulates Adhesion and Migration of Human Gingival Epithelial Cells Induced by IL-1β[J]. Journal of Oral Science Research, 2022, 38(6): 572-577.

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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

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