经典Wnt/β-catenin信号通路在朊病毒感染细胞模型SMB-S15中抑制的研究  

Remarkable impairment of Wnt/β-catenin signaling in prion infected cell model SMB-S15

作　　者：孙静[1] 王晶[1] 陈利娜[1] 杨晓东[1] 肖康[1] 陈操[1] 田婵[1] 石琦[1] 董小平[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所朊病毒病室传染病预防控制国家重点实验室,北京102206

出　　处：《中华实验和临床病毒学杂志》2016年第2期117-119,共3页Chinese Journal of Experimental and Clinical Virology

基　　金：国家自然科学基金项目（31270185,81301429,81572048）;传染病预防控制国家重点实验室基金（2012SKLIDl02,2015SKLID503）

摘　　要：目的 检测经典Wnt/β-catenin信号通路在朊病毒感染的细胞模型SMB-S15中的功能状态.方法 运用real time-PCR方法检测了朊病毒感染的细胞模型SMB-S15中,经典Wnt/β-catenin信号通路调控的靶基因mRNA表达水平;应用Western Blot方法检测稳定感染羊瘙痒因子Chandler株的小鼠神经细胞系SMB-S15中经典Wnt/β-catenin信号通路的相关调控蛋白及靶蛋白的表达.结果 与磷酸戊聚糖治愈的细胞对照SMB-PS相比,SMB-S15细胞中经典Wnt信号通路的靶基因cyclin D1、c-myc、survivin转录水平均有显著下调.经典Wnt信号通路的调控蛋白中,磷酸化p-catenin （Ser33,37,Thr41）表达明显上调,且其靶基因cyclin D1的蛋白表达明显下调,失活形式磷酸化糖原合成激酶（GSK-3β Ser9）表达明显下调.结论 在朊病毒感染的细胞模型SMB-S15中经典Wnt/β-catenin信号通路被抑制.Objective To investigate the functional state of Wnt/β-catenin signaling in prion infected cell model.Methods Evaluate alterations in the factors involved in the down-regulation of canonical Wnt signaling in prion infected cell line SMB-S15 by real time-PCR and Western blot.Results Compared with the control cell line SMB-PS,the transcriptions of Wnt/β-catenin signaling target genes cyclin D1 、c-myc、survivin were down-regulated in scrapie infected cell line SMB-S15 in the assays of real time-PCR.Western blots revealed that the levels of phosphor-β-catenin Ser33,37,Thr41 （p-β-catenin Ser33,37,Thr41） SMB-S15 cell line were much higher than that of SMB-PS cell,while that of cyclin D1,one of the target genes of Wnt signaling,was decreased.The level of phosphor-glycogen synthase kinase-3β （GSK-3β） Ser9 were markedly reduced,representing an enhanced GSK-3β activity in SMB-S15 cells.Conclusion Our data here strongly indicate an impairment of Wnt/β-catenin pathway in prion infected cell line SMB-S15.

关 键 词：朊病毒病 WNT/Β-CATENIN信号通路 GSK-3Β GSK-3β靶基因 

分 类 号：R742.9[医药卫生—神经病学与精神病学]