中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (30): 4767-4772.doi: 10.12307/2022.755

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

敲低环状RNA mmu_circ_0001775可提高小鼠骨髓间充质干细胞的成骨能力

董  艺1,单  帅1,刘佳林1,韩祥帧1,何惠宇1,2   

  1. 1新疆医科大学第一附属医院(附属口腔医院)口腔修复与种植科,新疆维吾尔自治区乌鲁木齐市   830054;2新疆维吾尔自治区口腔医学研究所,新疆维吾尔自治区乌鲁木齐市   830054
  • 收稿日期:2021-09-25 接受日期:2021-11-10 出版日期:2022-10-28 发布日期:2022-03-29
  • 通讯作者: 何惠宇,主任医师,教授,博士、硕士生导师,新疆医科大学第一附属医院(附属口腔医院)口腔修复与种植科,新疆维吾尔自治区乌鲁木齐市 830054;新疆维吾尔自治区口腔医学研究所,新疆维吾尔自治区乌鲁木齐市 830054
  • 作者简介:董艺,女,1995年生,山东省烟台市人,在读硕士,医师,主要从事口腔修复学及组织工程研究。
  • 基金资助:
    国家自然科学基金项目(81660177),项目负责人:何惠宇;自治区研究生创新项目(XJ2021G196),项目负责人:刘佳林

Circular RNA mmu_circ_0001775 knockdown improves the osteogenic ability of mouse bone marrow mesenchymal stem cells

Dong Yi1, Shan Shuai1, Liu Jialin1, Han Xiangzhen1, He Huiyu1, 2   

  1. 1Department of Prosthodontics and Implant, First Affiliated Hospital (Affiliated Stomatological Hospital) of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; 2Institute of Stomatology, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • Received:2021-09-25 Accepted:2021-11-10 Online:2022-10-28 Published:2022-03-29
  • Contact: He Huiyu, Chief physician, Professor, Doctoral and master’s supervisor, Department of Prosthodontics and Implant, First Affiliated Hospital (Affiliated Stomatological Hospital) of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; Institute of Stomatology, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • About author:Dong Yi, Master candidate, Physician, Department of Prosthodontics and Implant, First Affiliated Hospital (Affiliated Stomatological Hospital) of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • Supported by:
    National Natural Science Foundation of China, No. 81660177 (to HHY); Autonomous Region Postgraduate Innovation Project, No. XJ2021G196 (to LJL)

摘要:

文题释义:
环状RNA:是一类以共价键形成环形结构的非编码RNA分子,广泛分布于真核生物中,对基因的表达具有重要调控作用。
微小RNA:是一类由内源基因组成的非编码单链RNA分子,通过直接与靶基因位点结合,可以正向或负向靶向成骨作用来调节细胞的成骨分化,并相应地促进或抑制细胞成骨。
竞争性抑制:环状RNA含有丰富microRNA的结合位点,可以作为竞争性内源RNA在细胞中起到microRNA分子海绵的作用,消除microRNA对靶基因的抑制作用,提高靶基因的表达水平。

背景:近年来利用环状RNA(circRNA)调控干细胞分化成为新的研究热点,将circRNA负载于骨髓间充质干细胞研究其成骨性能,为研发骨缺损修复新型材料具有重要意义。
目的:探讨敲低circRNA mmu_circ_0001775对骨髓间充质干细胞成骨能力的影响。
方法:生物信息学分析工具推测miR-335-5p关系最为密切的上游circRNA为mmu_circ_0001775,将敲低mmu_circ_0001775的慢病毒、阴性对照病毒转染至骨髓间充质干细胞。qRT-PCR检测各组细胞成骨诱导7,14 d时Runt相关转录因子2、骨钙素、miR-335-5p及mmu_circ_0001775的表达水平,验证其成骨作用,Western blot检测骨形态发生蛋白2、骨桥蛋白的表达。
结果与结论:LV-mmu_circ_0001775-BMMSCs组各时间点Runt相关转录因子2、骨钙素表达高于LV-BMMSCs组(P < 0.01),骨形态发生蛋白2、骨桥蛋白相对表达量均明显高于LV-BMMSCs组(P < 0.05)。结果表明,敲低环状RNA mmu_circ_0001775有利于提高骨髓间充质干细胞的成骨能力。

https://orcid.org/0000-0002-8832-5189 (董艺)

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: circRNA, microRNA, 成骨分化, 颌骨缺损, 组织再生, 骨髓间充质干细胞, 分子海绵

Abstract: BACKGROUND: In recent years, the use of circular RNA (circRNA) to regulate stem cell differentiation has become a new research focus. Therefore, loading circRNA on bone marrow mesenchymal stem cells to study their osteogenic properties is of great significance for the development of new materials to repair bone defect.  
OBJECTIVE: To investigate the osteogenic effect of circRNA mmu_circ_0001775 knockdown on bone marrow mesenchymal stem cells.
METHODS:  Bioinformatics analysis tools speculated that the upstream circRNA most closely related to miR-335-5p was mmu_circ_0001775. Lentivirus and negative control virus knockdown mmu_circ_0001775 were transfected into bone marrow mesenchymal stem cells. qRT-PCR was used to detect the expression levels of Runt-related transcription factor 2, osteocalcin, miR-335-5p, and mmu_circ_0001775 genes at 7 and 14 days of osteogenic induction, respectively, to verify their osteogenic effects. Western blot assay was used to detect the expression levels of bone morphogenetic protein 2 and osteopontin.  
RESULTS AND CONCLUSION: The expression levels of Runt-related transcription factor 2 and osteocalcin at each time point in LV-mmu_circ_0001775-BMMSCs group were higher than those in LV-BMMSCs group (P < 0.01). The relative expression levels of bone morphogenetic protein 2 and osteopontin proteins were significantly higher in the LV-mmu_circ_0001775-BMMSCs group than those of LV-BMMSCs group (P < 0.05). The results showed that cirCRNA mmu_circ_0001775 knockdown could enhance the osteogenic ability of bone marrow mesenchymal stem cells.

Key words: circRNA, microRNA, osteogenic differentiation, jaw defect, tissue regeneration, bone marrow mesenchymal stem cells, molecular sponge

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