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

作　　者：郭静远[1] 王权[1] 温轩[1] 谭乔燕[1] 谢杨丽[1] 苏楠[1] 陈林[1] 

机构地区：[1]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,重庆400042

出　　处：《第三军医大学学报》2017年第10期946-953,共8页Journal of Third Military Medical University

基　　金：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)~~

摘　　要：目的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 noncoding RNA,LncRNA)Malat1的调控作用。方法采用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 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。Objective To investigate the role of fibroblast growth factor pathway in the regulation of long non-coding RNA（LncRNA） Malatl in chondrocytes. Methods Primary chondrocytes isolated from mice with tissue-specific knockout of fibroblast growth factor receptor 1 （ Fgfrl ） 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 Malatl in the cells was detected with qPCR. ATDC5 cells with small interfering RNA （siRNA） -mediated silencing of Malatl or Fgfrl gene and cells with Fgfrl overexpression were examined for the expression level of Malatl using qPCR, and the cell proliferation was assessed by cell counting. Results The proliferation of ATDC5 cells was suppressed after siRNA-mediated silencing of Malatl. Treatment with FGF2 significantly down-regulated the expression level of Malatl in ATDC5 cells, and this inhibitory effect was abolished by the application of the ERK pathway blocker U0126. Conditional knockout or interference of Fgfrl resulted in significant up-regulation of Malatl expression and abolished the inhibitory effect of FGF2 in ATDC5 cells, while overexpression of Fgfrl in ATDC5 cells caused significant down-regulation of Malatl expression. Conclusion In chondrocytes, FGF2 inhibits Malatl expression by activating the ERK pathway through Fgfrl suppressing the cell proliferation.

关 键 词：长链非编码RNA Malat1 成纤维细胞生长因子 软骨细胞 增殖 

分 类 号：R322.71[医药卫生—人体解剖和组织胚胎学] R329.28[医药卫生—基础医学]