Yang Longfei, Zhang Jinlong, Chen Jiajia, Jin Huricha, Liu Jian, Huang Shen, Cui Zhiming
Department of Spine Surgery, The Second Affiliated Hospital of Nantong University, Nantong University, Nantong, 226001, Jiangsu, People's Republic of China.
Neurochem Res. 2015 Sep;40(9):1966-75. doi: 10.1007/s11064-015-1692-0. Epub 2015 Aug 18.
CUG-binding protein 1, a member of the CELF (CUGBP and embryonic lethal abnormal vision-like factor) family of RNA-binding proteins, is shown to be multifunctional, regulating many posttranscriptional processes including alternative splicing, deadenylation, mRNA decay, and translation. Recently, CUGBP1 is found to represses p27 IRES activity and inhibits expression of endogenous p27 in cultured breast cancer cells. However, the roles of CUGBP1 in central nervous system injury remain unknown. In our study, we performed acute spinal cord injury (SCI) model in adult rats in order to research the expression changes of CUGBP1 in spinal cord. Western blot analysis showed a marked upregulation of CUGBP1 after SCI. Immunohistochemistry analysis revealed a wide distribution of CUGBP1 in the spinal cord. Double immunofluorescence staining indicated that CUGBP1 immunoreactivity was increased predominantly in neurons and astrocytes after SCI. Moreover, colocalization of CUGBP1/proliferating cell nuclear antigen (PCNA) was detected in GFAP positive cells. We also examined the expression profiles of p27, which was up-regulated after SCI. To further understand whether CUGBP1 plays a role in astrocyte proliferation, we applied LPS to induce astrocyte proliferation in vitro. Western blot analysis demonstrated that CUGBP1 expression was positively correlated with PCNA expression, and the p27 expression was negatively correlated with CUGBP1 expression following LPS stimulation. Our results suggest that CUGBP1 might be implicated in the pathophysiology of spinal cord after SCI.
CUG结合蛋白1是CELF(CUGBP和胚胎致死性异常视觉样因子)家族RNA结合蛋白的成员之一,具有多种功能,可调节包括可变剪接、去腺苷酸化、mRNA降解和翻译在内的许多转录后过程。最近,研究发现CUGBP1可抑制p27内部核糖体进入位点(IRES)活性,并抑制培养的乳腺癌细胞中内源性p27的表达。然而,CUGBP1在中枢神经系统损伤中的作用仍不清楚。在我们的研究中,我们在成年大鼠中建立了急性脊髓损伤(SCI)模型,以研究脊髓中CUGBP1的表达变化。蛋白质免疫印迹分析显示,SCI后CUGBP1显著上调。免疫组织化学分析显示CUGBP1在脊髓中广泛分布。双重免疫荧光染色表明,SCI后CUGBP1免疫反应性主要在神经元和星形胶质细胞中增加。此外,在GFAP阳性细胞中检测到CUGBP1与增殖细胞核抗原(PCNA)的共定位。我们还检测了p27的表达谱,其在SCI后上调。为了进一步了解CUGBP1是否在星形胶质细胞增殖中发挥作用,我们应用脂多糖(LPS)在体外诱导星形胶质细胞增殖。蛋白质免疫印迹分析表明,LPS刺激后,CUGBP1表达与PCNA表达呈正相关,而p27表达与CUGBP1表达呈负相关。我们的结果表明,CUGBP1可能参与了SCI后脊髓的病理生理过程。