阿尔茨海默病细胞模型中lncRNA RP11-543N12.1对CDH13表达的调控作用  

Regulatory effect of lncRNA RP11-543N12.1 on the expression of CDH13 in Alzheimer's disease cell model

作　　者：李蒙蒙[1] 王苗苗[1] 刁雪琴 田克立[1] 徐霞[1] 任桂杰[1] 

机构地区：[1]山东大学医学院生物化学与分子生物学系,山东济南250012

出　　处：《山东大学学报（医学版）》2017年第3期12-18,31,共8页Journal of Shandong University:Health Sciences

基　　金：山东省自然科学基金(ZR2015HM027);中国博士后科学基金(20090461219);山东省卫生厅资助项目(2014WS0405)

摘　　要：目的探讨阿尔茨海默病(AD)细胞模型中长链非编码RNA(lncRNA)RP11-543N12.1对CDH13的表达调控作用。方法采用基因芯片技术筛选AD相关差异表达lncRNA和mRNA,并通过Real-time PCR技术验证AD细胞模型中lncRNA RP11-543N12.1和CDH13 mRNA表达均增加,与芯片结果一致;然后将RP11-543N12.1-siRNA转入SH-SY5Y细胞,并加入β-淀粉样蛋白(Aβ25-35)作用48 h,接着MTT检测细胞存活率,Real-time PCR技术检测RP11-543N12.1和CDH13在RNA水平的变化,Western blotting检测CDH13蛋白表达,Hoechst33258染色检测细胞核的形态学变化,流式细胞术检测凋亡率。结果在未转染RP11-543N12.1-siRNA的SH-SY5Y细胞中,Aβ25-35处理48 h后,与正常细胞相比,细胞存活率降低(P<0.05),CDH13 RNA水平增加(P<0.01),CDH13蛋白活性形式表达量增加(P<0.01),细胞核出现凋亡的形态学变化,流式检测细胞凋亡率增加(P<0.01);转入RP11-543N12.1-siRNA后再用Aβ25-35处理48 h,细胞存活率、CDH13 RNA水平、蛋白活性形式表达量、细胞凋亡率均趋于正常水平,细胞核凋亡的形态学变化消失。结论 LncRNA RP11-543N12.1能够调节CDH13的表达,降低RP11-543N12.1的表达量能使CDH13表达下调。Objective To explore the regulatory function of long noncoding RNA（lncRNA） RP11-543N12.1 on the expression of CDH13 in Alzheimer＇s disease（AD） cell model.Methods Firstly,differently expressed lncRNAs and mRNAs were screened through microarray,and upregulated expressions of both RP11-543N12.1 and CDH13 were verified by real-time PCR.Secondly,SH-SY5Y cells were transfected with RP11-543N12.1-siRNA,and then Aβ25-35 was added to the transfected cells.After 48 h,the survival rate was detected with M TT;RNA expressions of RP11-543N12.1and CDH13 were detected with real-time PCR;protein expression of CDH13 was determined with Western blotting;morphological changes were observed with Hoechst33258 staining;cellular apoptosis rate was detected with flow cytometry.Results Compared with control SH-SY5 Y cells,cells treated with Aβ25-35 showed inhibited proliferation（P〈0.05）,upregulated RNA and protein expressions of CDH13（both P〈0.01）,and increased cellular apoptosis rate（P〈0.01）.Apoptotic nuclei were also observed.In cells transfected with RP11-543N12.1-siRNA and treated with Aβ25-35,cellular survival rate,RNA and protein expressions,and cellular apoptosis rate were all close to normal levels.Nuclei apoptosis disappeared.Conclusion LncRNA RP11-543N12.1 can regulate expression of CDH13,and knockdown of RP11-543N12.1 can downregulate the expression of CDH13.

关 键 词：阿尔茨海默病 SH-SY5Y细胞 LncRNA RP11-543N12.1 CDH13 表达调控 

分 类 号：R574[医药卫生—消化系统]