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成纤维细胞生长因子抑制长链非编码RNA Malat1的表达及对软骨细胞增殖的影响
Role of fibroblast growth factor pathway in regulating expression of long non-coding RNA Malat1 and proliferation of chondrocytes
in vitro - 第三军医大学学报 2017年39卷第10期 页码：946-953
- 作者机构：
400042 重庆，第三军医大学大坪医院野战外科研究所创伤实验室，骨代谢与修复中心，创伤、烧伤与复合伤国家重点实验室
- 作者简介：
[陈林，E-mail: linchen70@163.com Chen Lin, E-mail: linchen70@163.com
- 基金信息：国家重点基础研究发展计划(“973”计划，2014CB942904);国家自然科学基金面上项目(81471092)
DOI：10.16016/j.1000-5404.201611251
中图分类号： R322.71;R329.28;R394.3收稿：2016-11-30，
修回：2017-02-13，
纸质出版：2017-05-30
- 稿件说明：
移动端阅览
郭静远, 王权, 温轩, 等. 成纤维细胞生长因子抑制长链非编码RNA Malat1的表达及对软骨细胞增殖的影响[J]. 第三军医大学学报, 2017,39(10):946-953.
Jingyuan Guo, Quan Wang, Xuan Wen, et al. Role of fibroblast growth factor pathway in regulating expression of long non-coding RNA Malat1 and proliferation of chondrocytes
in vitro [J]. Journal of Third Military Medical University, 2017, 39(10): 946-953.郭静远, 王权, 温轩, 等. 成纤维细胞生长因子抑制长链非编码RNA Malat1的表达及对软骨细胞增殖的影响[J]. 第三军医大学学报, 2017,39(10):946-953. DOI： 10.16016/j.1000-5404.201611251.
Jingyuan Guo, Quan Wang, Xuan Wen, et al. Role of fibroblast growth factor pathway in regulating expression of long non-coding RNA Malat1 and proliferation of chondrocytes
in vitro [J]. Journal of Third Military Medical University, 2017, 39(10): 946-953. DOI： 10.16016/j.1000-5404.201611251.
摘要
目的
2
探讨在软骨细胞模型中成纤维细胞生长因子通路对长链非编码RNA(long non-coding RNA，LncRNA)Malat1的调控作用。
方法
2
采用Ⅰ型成纤维细胞生长因子受体(fibroblast growth factor receptor 1，Fgfr1) 组织特异性敲除小鼠和Ⅲ型成纤维细胞生长因子受体(fibroblast growth factor receptor 3，Fgfr3) 敲除小鼠的原代软骨，以及利用软骨细胞系培养充当软骨细胞模型，成纤维细胞生长因子2(fibroblast growth factor 2，FGF2) 及下游细胞外调节蛋白激酶(extracellular regulated protein kinases，ERK)通路阻断剂U0126处理细胞，利用荧光定量PCR检测Malat1的表达变化。在ATDC5细胞模型中通过siRNA转染干扰Malat1或Fgfr1，质粒转染过表达Fgfr1，利用荧光定量PCR检测Malat1的表达，细胞计数检测细胞增殖变化。
结果
2
Malat1被干扰24、36、48、60 h后，ATDC5细胞增殖速度显著减慢(
P
<
0.05)，FGF2处理ATDC5细胞24 h后Malat1表达显著下降(
P
<
0.05)，且ERK通路抑制剂U0126处理细胞后FGF2对Malat1的抑制作用消失。Fgfr1条件性敲除或si-Fgfr1干扰后Malat1表达显著升高(
P
<
0.05)，且FGF2对Malat1的抑制作用消失，Fgfr1在ATDC5细胞中过表达后Malat1表达与对照组相比显著降低(
P
<
0.05)。
结论
2
软骨细胞中FGF2可能通过Fgfr1激活下游ERK通路，抑制了Malat1的表达，从而影响软骨细胞增殖。
Abstract
Objective
2
To investigate the role of fibroblast growth factor pathway in the regulation of long non-coding RNA(LncRNA) Malat1 in chondrocytes.
Methods
2
Primary chondrocytes isolated from mice with tissue-specific knockout of fibroblast growth factor receptor 1 (Fgfr1) and Fgfr3 and
in vitro
cultured chondrocyte lines were treated with fibroblast growth factor 2 (FGF2) and U0126
a blocker of the downstream extracellular regulated protein kinases (ERK)
and the expression level of Malat1 in the cells was detected with qPCR. ATDC5 cells with small interfering RNA (siRNA)-mediated silencing of Malat1 or Fgfr1 gene and cells with Fgfr1 overexpression were examined for the expression level of Malat1 using qPCR
and the cell proliferation was assessed by cell counting.
Results
2
The proliferation of ATDC5 cells was suppressed after siRNA-mediated silencing of Malat1. Treatment with FGF2 significantly down-regulated the expression level of Malat1 in ATDC5 cells
and this inhibitory effect was abolished by the application of the ERK pathway blocker U0126. Conditional knockout or interference of Fgfr1 resulted in significant up-regulation of Malat1 expression and abolished the inhibitory effect of FGF2 in ATDC5 cells
while overexpression of Fgfr1 in ATDC5 cells caused significant down-regulation of Malat1 expression.
Conclusion
2
In chondrocytes
FGF2 inhibits Malat1 expression by activating the ERK pathway through Fgfr1 suppressing the cell proliferation.
关键词
Keywords
references
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