中国组织工程研究 ›› 2024, Vol. 28 ›› Issue (13): 2017-2023.doi: 10.12307/2024.166

• 脐带脐血干细胞 umbilical cord blood stem cells • 上一篇    下一篇

miR-26b对人脱落乳牙牙髓干细胞及人脐带间充质干细胞增殖、迁移和成骨分化的影响

袁媛园1,潘  露1,周绍兰1,梁  燕1,许键炜2,徐  文1   

  1. 1贵州医科大学附属口腔医院,贵州省贵阳市   550004;2贵州医科大学组织工程与干细胞实验中心,贵州省贵阳市   550004
  • 收稿日期:2023-04-18 接受日期:2023-06-08 出版日期:2024-05-08 发布日期:2023-08-28
  • 作者简介:袁媛园,女,1984年生,湖南省长沙市人,壮族,2009年贵州医科大学(原贵阳医学院)毕业,硕士,副主任医师,主要从事牙体牙髓病学及牙髓干细胞研究工作。
  • 基金资助:
    贵州省卫生健康委科学技术基金项目(gzwjkj2021-1-353),项目负责人:袁媛园;贵州医科大学附属口腔医院学科项目(KQYY-2021-2),项目负责人:袁媛园

Effect of miR-26b on proliferation, migration and osteogenic differentiation of stem cells from human exfoliated deciduous teeth and human umbilical cord mesenchymal stem cells

Yuan Yuanyuan1, Pan Lu1, Zhou Shaolan1, Liang Yan1, Xu Jianwei2, Xu Wen1   

  1. 1Affiliated Stomatological Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China; 2Tissue Engineering and Stem Cell Experiment Center of Guizhou Medical University, Guiyang 550004, Guizhou Province, China
  • Received:2023-04-18 Accepted:2023-06-08 Online:2024-05-08 Published:2023-08-28
  • About author:Yuan Yuanyuan, Master, Associate chief physician, Affiliated Stomatological Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China
  • Supported by:
    cience and Technology Fund Project of Guizhou Provincial Health Commission, No. gzwjkj2021-1-353 (to YYY); Discipline Project of Dental Hospital Affiliated to Guizhou Medical University, No. KQYY-2021-2 (to YYY)

摘要:


文题释义:

microRNAs(miRNAs):是存在于真核生物中的一组单链非编码RNA,长度约为22 nt,通过与特定转录物的3’非翻译区相互作用,促使其降解或抑制其翻译,来达到调节基因表达的目的,在细胞的生理活动中发挥重要作用。
细胞转染:是指细胞经一定处理后,细胞膜的通透性提高,使外源DNA或RNA片段能更容易进入细胞内部,从而获得新的表型的过程,可分为瞬时转染和稳定转染。


背景:microRNA-26b(miR-26b)对干细胞的多种功能具有重要的调节作用,但其对人脱落乳牙牙髓干细胞和人脐带间充质干细胞生物学特性的影响尚不清楚。

目的:探讨miR-26b对人脱落乳牙牙髓干细胞和人脐带间充质干细胞增殖、迁移和成骨分化的影响。
方法:培养及鉴定人脱落乳牙牙髓干细胞和人脐带间充质干细胞;用miR-26 mimics(实验组)和miRNAs mimics对照(对照组)转染2种细胞,构建过表达模型用于后续实验。CCK-8法检测过表达miR-26b细胞的增殖能力;Transwell和划痕实验分析过表达miR-26b细胞的迁移能力;RT-qPCR法检测过表达miR-26b细胞成骨诱导后成骨标志物的表达。

结果与结论:①转染miR-26b模拟物可提高2种细胞中miR-26b表达量,促进人脱落乳牙牙髓干细胞增殖,对人脐带间充质干细胞的扩增无明显影响;②相较于对照组,2种细胞过表达miR-26b后迁移能力增强;③miR-26b在2种细胞成骨分化过程中表达水平降低;④相较于对照组,2种细胞过表达miR-26b后,成骨相关基因骨钙素、骨桥素、碱性磷酸酶、Ⅰ型胶原蛋白mRNA水平均下调。结果表明,过表达miR-26b能促进人脱落乳牙牙髓干细胞增殖、迁移,抑制其成骨分化;也促进人脐带间充质干细胞迁移,抑制其成骨分化,但对其增殖无明显影响。

https://orcid.org/0009-0002-0806-3498 (袁媛园) 

中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程

关键词: miR-26b, 人脱落乳牙牙髓干细胞, 人脐带间充质干细胞, 迁移, 增殖, 成骨分化

Abstract: BACKGROUND: microRNA-26b (miR-26b) plays an important regulatory role in a variety of stem cell functions, but its effects on the biological properties of stem cells from human exfoliated deciduous teeth and human umbilical cord mesenchymal stem cells are unknown.
OBJECTIVE: To investigate the effects of miR-26b on the proliferation, migration and osteogenic differentiation of stem cells from human exfoliated deciduous teeth and human umbilical cord mesenchymal stem cells. 
METHODS: Stem cells from human exfoliated deciduous teeth and human umbilical cord mesenchymal stem cells were cultured and identified. miR-26 mimics (experimental group) and miRNAs mimics control (control group) were used to transfect above mentioned two kinds of cells and construct overexpressed models for subsequent experiments. CCK-8 assay was applied to detect the proliferation ability of overexpressed miR-26b cells. Transwell and scratch assay were employed to analyze the migration ability of overexpressed miR-26b cells. RT-qPCR was utilized to examine the expression of osteogenic markers after osteogenic induction of overexpressed miR-26b cells.
RESULTS AND CONCLUSION: (1) Transfection of miR-26b mimics increased miR-26b expression in the two kinds of cells and promoted the proliferation of stem cells from human exfoliated deciduous teeth, with no significant effect on the amplification of human umbilical cord mesenchymal stem cells. (2) Compared with the control group, the migration ability was enhanced after two types of cells overexpressing miR-26b. (3) miR-26b expression decreased during osteogenic differentiation of the two kinds of cells. (4) Compared with the control group, the levels of osteogenesis-related genes osteocalcin, osteopontin, alkaline phosphatase, and human type I collagen mRNA were downregulated after overexpression of miR-26b in the two kinds of cells. The results showed that overexpression of miR-26b promoted the proliferation and migration of stem cells from human exfoliated deciduous teeth and inhibited their osteogenic differentiation; it promoted the migration of human umbilical cord mesenchymal stem cells and inhibited their osteogenic differentiation, but had no significant effects on their proliferation.

Key words: miR-26b, stem cells from human exfoliated deciduous teeth, human umbilical cord mesenchymal stem cell, migration, proliferation, osteogenic differentiation

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