中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (25): 4038-4043.doi: 10.3969/j.issn.2095-4344.2017.25.017

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

长链非编码RNA DANCR促进滑膜间充质干细胞向软骨细胞的增殖与分化

杨 超1,张 雷2,周利武2,赵建宁2   

  1. 1南京大学医学院临床学院(解放军南京军区南京总医院)骨科,江苏省南京市 210000;2解放军南京军区南京总医院骨科,江苏省南京市  210002
  • 修回日期:2017-04-06 出版日期:2017-09-08 发布日期:2017-10-09
  • 通讯作者: 张雷,博士后,主治医师,解放军南京军区南京总医院骨科,江苏省南京市 210002; 赵建宁,主任医师,解放军南京军区南京总医院骨科,江苏省南京市 210002
  • 作者简介:杨超,男,1990年生,南京大学医学院在读硕士,主要从事关节软骨修复的研究。
  • 基金资助:

    江苏省临床医学科技专项资助(BL2012002);中国博士后基金(2016M592956);南京总医院院管课题(2016003)

Long non-coding RNA DANCR enhances chondrogenic differentiation and proliferation of human synovium-derived mesenchymal stem cells  

Yang Chao1, Zhang Lei2, Zhou Li-wu2, Zhao Jian-ning2   

  1. 1Department of Orthopedics, Clinical College of Medical School of Nanjing University, Nanjing 210000, Jiangsu Province, China; 2Department of Orthopedics, Nanjing General Hospital of Nanjing Military Command, Nanjing 210002, Jiangsu Province, China
  • Revised:2017-04-06 Online:2017-09-08 Published:2017-10-09
  • Contact: Zhang Lei, M.D., Attending physician, Department of Orthopedics, Nanjing General Hospital of Nanjing Military Command, Nanjing 210002, Jiangsu Province, China; Zhao Jian-ning, Chief physician, Department of Orthopedics, Nanjing General Hospital of Nanjing Military Command, Nanjing 210002, Jiangsu Province, China
  • About author:Yang Chao, Studying for master’s degree, Department of Orthopedics, Clinical College of Medical School of Nanjing University, Nanjing 210000, Jiangsu Province, China
  • Supported by:

    the Clinical Medical Research Foundation of Jiangsu Province of China, No. BL2012002; Postdoctoral Science Foundation of China, No. 2016M592956; the Funding Project of Nanjing General Hospital of Nanjing Military Command in China, No. 2016003

摘要:

文章快速阅读:

文题释义:
长链非编码RNA DANCR:
是一种在肿瘤中被发现的高表达的长链非编码RNA,通过生物信息学发现长链非编码RNA DANCR上游启动子存在 NF-kB和p-STAT3 的转录结合位点,其与CTNNB1 mRNA的表达密切相关,而后者直接编码β-catenin,而Wnt/β-catenin通路同样也是滑膜间充质干细胞向软骨细胞增殖与分化的重要调控通路。
长链非编码RNA DANCR与干细胞分化:目前的研究主要集中在肝细胞癌、大肠癌等肿瘤方面,但已有研究表明,长链非编码RNA DANCR在牙组织干细胞多向分化及滑膜间充质干细胞向软骨细胞增殖与分化中起着积极的调控作用,其可促进牙组织干细胞向成骨、成脂、神经等多方向分化。另有研究表明,长链非编码RNA MIAT的敲除可促进人脂肪来源的间充质干细胞向成骨细胞分化。

 

摘要
背景:
多项研究表明,长链非编码RNA DANCR可通过Wnt/β-catenin信号通路在多种病理生理过程中发挥重要作用。
目的:探究长链非编码RNA DANCR在滑膜间充质干细胞向软骨细胞增殖与分化过程中的作用。
方法:取第3代人滑膜间充质干细胞,分别以pcDNA3.1-GP质粒和pcDNA3.1-GP(DANCR Homo)质粒转染滑膜间充质干细胞,分别设为对照组和实验组,培养7 d内,检测两组细胞增殖曲线;培养14 d后,组织学观察两组细胞成软骨能力;qRT-PCR检测两组成软骨分化后细胞的软骨特异性基因表达。
结果与结论:①细胞增殖曲线:两组细胞增殖曲线大致呈“S”型,一二天内细胞处于相对停滞期,从第3天开始细胞进入快速增殖时期,实验组细胞增殖速度明显快于对照组;②组织学观察:两组细胞外基质均呈蓝色,符合软骨细胞特征,实验组细胞外基质蓝染强于对照组;③qRT-PCR检测:成软骨诱导14 d后,实验组软骨特异性基因Ⅱ型胶原、蛋白多糖、Sox9基因表达显著高于对照组(P < 0.05);④结果表明:长链非编码RNA DANCR可促进滑膜间充质干细胞向软骨细胞的增殖与分化,并加强软骨特异性基因的表达。

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
ORCID: 0000-0002-5975-6342(张雷)

关键词: 干细胞, 分化, 滑膜间充质干细胞, 长链非编码RNA, DANCR, 细胞增殖, 细胞分化

Abstract:

BACKGROUND: A number of studies have shown that long non-coding RNA DANCR can play an important role in various pathophysiological processes through Wnt/β-catenin signaling pathway.
OBJECTIVE: To explore the effect of long non-coding RNA DANCR on the proliferation and chondrogenesis of synovium-derived mesenchymal stem cells.
METHODS: Passage 3 synovium-derived mesenchymal stem cells were obtained and transfected with pcDNA3.1-GP (control) and pcDNA3.1-GP(DANCR Homo) (experimental). Cell viability was estimated at 1, 2, 3, 4, 5, 6 and 7 days after DANCR transfection. The passage 3 cells were cultured in the chondriogenic medium for 14 days. And the chondrogenesis potential of cells was examined by toluidine blue staining. The chondrogenic-specific marker genes Aggrecan, Type II collagen (Col2) and Sox9 were determined by Real-time PCR.
RESULTS AND CONCLUSION: The synovium-derived mesenchymal stem cells exhibited “S”-shaped curves in the two groups, with cell arrest at 1-2 days and rapid proliferation beginning at 3 days. Cell counting kit-8 assay and toluidine blue staining showed overexpressing DANCR significantly promoted proliferation in synovium-derived mesenchymal stem cells. The aggregates from synovium-derived mesenchymal stem cells in the experimental group had a greater amount of toluidine blue staining than the control group. In addition, we detected the higher expression of chondrogenic specific marker genes, such as Col2, Sox9 and Aggrecan, in the experimental group than the control group at 14 days after chondrogenic induction (P < 0.05). These results demonstrate that long non-coding RNA DANCR could enhance chondrogenic differentiation and proliferation of human synovium-derived mesenchymal stem cells and increase the expression of chondrogenic specific marker genes.

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

Key words: Stem Cells, Mesenchymal Stem Cells, Cell Proliferation, Cell Differentiation, Tissue Engineering

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