中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (20): 3223-3229.doi: 10.3969/j.issn.2095-4344.2014.20.020

• 骨组织构建 bone tissue construction • 上一篇    下一篇

骨形态发生蛋白2诱导C3H10T1/2细胞成骨分化中长链非编码RNA的作用

程  晨1,高  艳1,李  静1,潘秋辉2   

  1. 同济大学附属第十人民医院,1检验科,2中心实验室,上海市  200072
  • 收稿日期:2014-02-28 出版日期:2014-05-14 发布日期:2014-05-14
  • 通讯作者: 潘秋辉,同济大学附属第十人民医院中心实验室,上海市 200072
  • 作者简介:程晨,女,1988年生,甘肃省嘉峪关市人,汉族,同济大学在读硕士,主要从事肿瘤分子生物学方面的研究。
  • 基金资助:

    国家自然科学基金资助项目(81171778)

Role of long non-coding RNA in osteoblast differentiation of C3H10T1/2 cells induced by bone morphogenetic protein-2

Cheng Chen1, Gao Yan1, Li Jing1, Pan Qiu-hui2   

  1. 1Department of Medical Laboratory, Tenth People’s Hospital, Tongji University, Shanghai 200072, China; 2Central Laboratory, Tenth People’s Hospital, Tongji University, Shanghai 200072, China
  • Received:2014-02-28 Online:2014-05-14 Published:2014-05-14
  • Contact: Pan Qiu-hui, Central Laboratory, Tenth People’s Hospital, Tongji University, Shanghai 200072, China
  • About author:Cheng Chen, Studying for master’s degree, Department of Medical Laboratory, Tenth People’s Hospital, Tongji University, Shanghai 200072, China
  • Supported by:

    Funding: the National Natural Science Foundation of China, No. 81171778

摘要:

背景:人类长链非编码RNA是现今的研究热点,已有研究报道其在肿瘤发生中的作用机制,但其在成骨分化方面的机制并不明确。
目的:观察人类长链非编码RNA在经骨形态发生蛋白2诱导小鼠C3H10T1/2细胞成骨分化中的作用,并探讨其分子机制。
方法:首先进行碱性磷酸酶染色和成骨指标基因检测。对C3H10T1/2细胞在骨形态发生蛋白2诱导下的成骨分化过程长链非编码RNA表达变化进行芯片分析。采用高通量测序比较骨形态发生蛋白2诱导组和未经骨形态发生蛋白2诱导组的表达变化,筛选出表达下降的基因。过表达相应长链非编码RNA后观察对骨形态发生蛋白2诱导成骨分化的影响。
结果与结论:骨形态发生蛋白2诱导C3H10T1/2细胞导致碱性磷酸酶活性增加。骨形态发生蛋白2诱导72 h后,碱性磷酸酶、Id1、骨钙素、Runx2、sp7表达上升(P < 0.05)。未诱导C3H10T1/2细胞与骨形态发生蛋白2诱导细胞芯片杂交后结果比较,下降达1.5倍的长链非编码RNAs有24条,其中只有AK035085有内含子。与未过表达AK035085的对照组相比,骨形态发生蛋白2诱导72 h后AK035085过表达的C3H10T1/2细胞碱性磷酸酶、Id1、骨钙素、Runx2、sp7表达均下降(P < 0.05)。提示骨形态发生蛋白2可刺激C3H10T1/2细胞发生成骨分化,AK035085可能对C3H10T1/2细胞的成骨分化存在抑制作用。



中国组织工程研究
杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程


全文链接:

关键词: 组织构建, 骨组织工程, 长链非编码RNA, C3H10T1/2细胞, 骨形态发生蛋白2, 成骨分化, 国家自然科学基金

Abstract:

BACKGROUND: Long non-coding RNAs (lncRNAs) have became the hot topic in current studies and play an important role in the tumorigenesis. However, lncRNAs involved in the osteoblast differentiation remain poorly reported.
OBJECTIVE: To investigate the role of human LncRNAs in osteogenic differentiation of C3H10T1/2 cells induced by bone morphogenetic protein-2 and explore action mechanism.
METHODS: The induction of bone morphogenetic protein-2 was validated by alkaline phosphatase staining and the expression of corresponding genes was detected. The lncRNA expression profile was analyzed using the Arraystar lncRNA array in C3H10T1/2 MSCs undergoing early osteoblast differentiation. The expression with or without bone morphogenetic protein-2 induction was compared with high-flux sequencing, and the down-regulated genes were screened. The effect of lncRNA overexpression on osteogenic differentiation of C3H10T1/2 cells induced by bone morphogenetic protein-2 was observed.
RESULTS AND CONCLUSION: The bone morphogenetic protein-2 induced C3H10T1/2 cells led to increased alkaline phosphatase activity. After 72 hours of bone morphogenetic protein-2 induction, alkaline phosphatase,
Id1,osteocalcin, Runx2, sp7 expression were increased (P < 0.05). There were 24 down-regulated lncRNAs identified between bone morphogenetic protein-2 treated and untreated groups, the decrease of expression was 1.5 folds, and among them, only AK035085 contained intron. Compared with control group with no AK03508 expression, over-expression lncRNA AK035085 decreased the expression of alkaline phosphatase, Id1, osteocalcin, Runx2, sp7   (P < 0.05). Experimental findings indicate that bone morphogenetic protein-2 induces osteogenic differentiation of C3H10T1/2 cells and AK035085 inhibits the osteogenic differentiation.



中国组织工程研究
杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程


全文链接:

Key words:  bone morphogenetic proteins, mesenchymal stem cells, tumor, alkaline phosphatase, osteoblasts

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