MicroRNA-9靶向BACE1参与Alzheimer病发生发展的机制研究
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摘要
目的探讨microRNA-9(miR-9)靶向BACE1参与Alzheimer病(AD)发生发展的机制。方法选取2013年5月-2015年5月解放军第161医院收治的AD患者23例,以血管性痴呆(Vascular dementia,VD)21例和颅内感染患者(intracranial infection,ICI)20例为对照组,比较3组患者脑脊液中miR-9、BACE1和APP的表达水平;然后用荧光素酶报告系统验证miR-9与BACE1之间的靶向关系;最后将miR-9 mimics转染入SH-SY5Y细胞,qRT-PCR和Western Bolt检测BACE1和APP的mRNA和蛋白表达水平。结果与VD和ICI组比较,AD患者脑脊液中miR-9表达水平下降,BACE1和APP表达水平增高,AD患者脑脊液中miR-9表达水平与BACE1表达水平呈负相关;BACE1为miR-9的功能靶基因;miR-9可靶向BACE1参与调控SH-SY5Y细胞APP表达。结论 miR-9可靶向BACE1参与AD的发生发展。
Objective To investigate the effects of microRNA-9(miR-9)Via targeting Beta-site Amyloid precursor protein Cleaving Enzyme(BACE1)on Alzheimer's disease(AD).Methods 23 patients with AD(AD group)and21 patients with Vascular dementia(VD)(VD group)(the control group),20 patients with intracranial infection(ICI)(ICI group)(the control group)were enrolled in our hospital from May 2013 to May 2015.The expression levels of miR-9,BACE1 and APP in CSF of AD、VD and ICI were detected and compared in three groups.Then wild-type and mutant luciferase reporter plasmids were constructed according to the predicted miR-9 binding site on the 3'-UTR of the BACE1 mRNA and transfected into human embryonic kidney 293 tool cells(HEK293T)to determine the targeting relationship of miR-9 and BACE1.The regulation effects of miR-9 on the BACE1 and APP expression level in human heuroblastoma SH-SY5Y cells were determined.Results Compared with VD group and ICI group,the expression levels of miR-9 in CSF of AD group,the expression levels of BACE1 and APP in CSF of AD group were increased,the expression level of miR-9 was negatively relation with the expression level of BACE1 in CSF of AD.And luciferase reporter assay demonstrated that miR-9 was shown to specifically bind to the 3'-UTR of BACE1.Further miR-9-mimic SH-SY5 Y cells showed significantly lower BACE1 mRNA and protein expression levels and APP accumulation than those of matching negative control and non-transfected cells.Conclusion MiR-9 was down-regulated in an association with the up-regulation of BACE1 and APP accumulation in AD CSF,and it could reduce the BACE1 expression level,and APP accumulation in vitro by targeting BACE1.These finding suggested that miR-9 played an important role in the process of AD via targeting BACE1 and might be an new therapeutic molecular target for AD.
Objective To investigate the effects of microRNA-9(miR-9)Via targeting Beta-site Amyloid precursor protein Cleaving Enzyme(BACE1)on Alzheimer's disease(AD).Methods 23 patients with AD(AD group)and21 patients with Vascular dementia(VD)(VD group)(the control group),20 patients with intracranial infection(ICI)(ICI group)(the control group)were enrolled in our hospital from May 2013 to May 2015.The expression levels of miR-9,BACE1 and APP in CSF of AD、VD and ICI were detected and compared in three groups.Then wild-type and mutant luciferase reporter plasmids were constructed according to the predicted miR-9 binding site on the 3'-UTR of the BACE1 mRNA and transfected into human embryonic kidney 293 tool cells(HEK293T)to determine the targeting relationship of miR-9 and BACE1.The regulation effects of miR-9 on the BACE1 and APP expression level in human heuroblastoma SH-SY5Y cells were determined.Results Compared with VD group and ICI group,the expression levels of miR-9 in CSF of AD group,the expression levels of BACE1 and APP in CSF of AD group were increased,the expression level of miR-9 was negatively relation with the expression level of BACE1 in CSF of AD.And luciferase reporter assay demonstrated that miR-9 was shown to specifically bind to the 3'-UTR of BACE1.Further miR-9-mimic SH-SY5 Y cells showed significantly lower BACE1 mRNA and protein expression levels and APP accumulation than those of matching negative control and non-transfected cells.Conclusion MiR-9 was down-regulated in an association with the up-regulation of BACE1 and APP accumulation in AD CSF,and it could reduce the BACE1 expression level,and APP accumulation in vitro by targeting BACE1.These finding suggested that miR-9 played an important role in the process of AD via targeting BACE1 and might be an new therapeutic molecular target for AD.
引文
[1]Bhattacharjee S,Zhao YH,Hill JM,et al.Aluminum and its potential contribution to Alzheimer's disease(AD)[J].Frontiers in Aging Neuroscience,2014,6(4):62.
[2]Cheng L,Doecke JD,Sharples RA,et al.Prognostic serum miRNA biomarkers associated with Alzheimer's disease shows concordance with neuropsychological and neuroimaging assessment[J].Molecular Psychiatry,2015,20(10):1188-1196.
[3]Danborg PB,Simonsen AH,Waldemar G,et al.The potential of microRNAs as biofluid markers of neurodegenerative diseases--a systematic review[J].Biomarkers,2014,19(4):259-268.
[4]Denk J,Boelmans K,Siegismund C,et al.MicroRNA profiling of CSF reveals potential biomarkers to detect alzheimer's disease[J].PLOS One,2015,10(5):e0126423.
[5]Serpente M,Fenoglio C,Villa C,et al.Circulating miRNAs as potential biomarkers in Alzheimer's disease[J].Journal of Alzheimers Disease,2014,41(2):S50.
[6]Deng Yan-yao,Din Yu,Hou D.Research status of the regulation of miRNA on BACE1[J].International Journal of Neuroscience,2014,124(7):474-477.
[7]Liu Huan,Tian Tian,Qin Shan-chun,et al.Folic acid deficiency enhances abeta accumulation in APP/PS1 mice brain and decreases amyloid-associated miRNAs expression[J].Journal of Nutritional Biochemistry,2015,26(12):1502-1508.
[8]Yang Guo-shuai,Song Yan-min,Zhou Xiao-yan,et al.MicroR-NA-29c targets beta-site amyloid precursor protein-cleaving enzyme 1 and has a neuroprotective role in vitro and in vivo[J].Molecular Medicine Reports,2015,12(2):3081-3088.
[9]Hill JM,Clement C,Pogue AI,et al.Pathogenic microbes,the microbiome,and Alzheimer's disease(AD)[J].Frontiers in Aging Neuroscience,2014,6(6):127.
[10]Zhao Yu-hai,Pogue AI,Lukiw WJ.MicroRNA(miRNA)signaling in the human CNS in sporadic alzheimer's disease(AD)-Novel and unique pathological features[J].International Journal of Molecular Sciences,2015,16(12):30105-30116.
[11]Zhao Yan-xin,Tan Wei,Sheng Wen-hua,et al.Identification of biomarkers associated with alzheimer's disease by bioinformatics analysis[J].American Journal of Alzheimers Disease and Other Dementias,2016,31(2):163-168.
[12]Chang Fei,Zhang Lin-hong,Xu Wu-ping,et al.microRNA-9attenuates amyloid beta-induced synaptotoxicity by targeting Calcium/calmodulin-dependent protein kinase kinase 2[J].Molecular Medicine Reports,2014,9(5):1917-1922.
[13]Kong Y,Wu J,Yuan L.MicroRNA expression analysis of adult-onset Drosophila Alzheimer's disease model[J].Current Alzheimer Research,2014,11(9):882-891.
[14]Cai L,Cai X.Up-regulation of miR-9 expression predicate advanced clinicopathological features and poor prognosis in patients with hepatocellular carcinoma[J].Diagn Pathol,2014,9(12):1000.
[15]Higashi T,Hayashi H,Ishimoto T,et al.miR-9-3p plays a tumour-suppressor role by targeting TAZ(WWTR1)in hepatocellular carcinoma cells[J].British Journal of Cancer,2015,113(2):252-258.
[16]Zhang Jiang-bo,Cheng Jin,Zeng Zhen-zhen,et al.Comprehensive profiling of novel microRNA-9 targets and a tumor suppressor role of microRNA-9 via targeting IGF2BP1 in hepatocellular carcinoma[J].Oncotarget,2015,6(39):42040-42052.
[17]Drakaki A,Hatziapostolou M,Polytarchou C,et al.Functional microRNA high throughput screening reveals miR-9 as a central regulator of liver oncogenesis by affecting the PPARA-CDH1 pathway[J].BMC Cancer,2015,15(7):542.
[18]Sun Zhao,Han Qin,Zhou Na,et al.MicroRNA-9 enhances migration and invasion through KLF17 in hepatocellular carcinoma[J].Molecular Oncology,2013,7(5):884-894.
[19]Liu Q-y,Chang M-n,Lei J-x,et al.Identification of microRNAs involved in Alzheimer's progression using a rabbit model of the disease[J].American Journal of Neurodegenerative Disease,2014,3(1):33-44.
[20]Romano GL,Platania CB,Forte S,et al.MicroRNA target prediction in glaucoma[J].Prog Brain Res,2015,220(6):217-240.
[21]Kropivsek K,Pickford J,Carter DA.Postnatal dynamics of Zeb2 expression in rat brain:analysis of novel 3'UTR sequence reveals a miR-9interacting site[J].Journal of Molecular Neuroscience,2014,52(1):138-147.
[22]Sim SE,Lim CS,Kim JI,et al.The Brain-Enriched MicroRNAmiR-9-3p regulates synaptic plasticity and memory[J].Journal of Neuroscience,2016,36(33):8641-8652.
[23]Liu Zhen,Wang Cun-fu,Wang Xiao,et al.Therapeutic effects of transplantation of As-MiR-937-Expressing mesenchymal stem cells in murine model of alzheimer's disease[J].Cellular Physiology and Biochemistry,2015,37(1):321-330.
[24]Jung HJ,Coffinier C,Choe Y,et al.Regulation of prelamin Abut not lamin C by miR-9,a brain-specific microRNA[J].Proceedings of the National Academy of Sciences of the United States of America,2012,109(7):E423-E431.
[2]Cheng L,Doecke JD,Sharples RA,et al.Prognostic serum miRNA biomarkers associated with Alzheimer's disease shows concordance with neuropsychological and neuroimaging assessment[J].Molecular Psychiatry,2015,20(10):1188-1196.
[3]Danborg PB,Simonsen AH,Waldemar G,et al.The potential of microRNAs as biofluid markers of neurodegenerative diseases--a systematic review[J].Biomarkers,2014,19(4):259-268.
[4]Denk J,Boelmans K,Siegismund C,et al.MicroRNA profiling of CSF reveals potential biomarkers to detect alzheimer's disease[J].PLOS One,2015,10(5):e0126423.
[5]Serpente M,Fenoglio C,Villa C,et al.Circulating miRNAs as potential biomarkers in Alzheimer's disease[J].Journal of Alzheimers Disease,2014,41(2):S50.
[6]Deng Yan-yao,Din Yu,Hou D.Research status of the regulation of miRNA on BACE1[J].International Journal of Neuroscience,2014,124(7):474-477.
[7]Liu Huan,Tian Tian,Qin Shan-chun,et al.Folic acid deficiency enhances abeta accumulation in APP/PS1 mice brain and decreases amyloid-associated miRNAs expression[J].Journal of Nutritional Biochemistry,2015,26(12):1502-1508.
[8]Yang Guo-shuai,Song Yan-min,Zhou Xiao-yan,et al.MicroR-NA-29c targets beta-site amyloid precursor protein-cleaving enzyme 1 and has a neuroprotective role in vitro and in vivo[J].Molecular Medicine Reports,2015,12(2):3081-3088.
[9]Hill JM,Clement C,Pogue AI,et al.Pathogenic microbes,the microbiome,and Alzheimer's disease(AD)[J].Frontiers in Aging Neuroscience,2014,6(6):127.
[10]Zhao Yu-hai,Pogue AI,Lukiw WJ.MicroRNA(miRNA)signaling in the human CNS in sporadic alzheimer's disease(AD)-Novel and unique pathological features[J].International Journal of Molecular Sciences,2015,16(12):30105-30116.
[11]Zhao Yan-xin,Tan Wei,Sheng Wen-hua,et al.Identification of biomarkers associated with alzheimer's disease by bioinformatics analysis[J].American Journal of Alzheimers Disease and Other Dementias,2016,31(2):163-168.
[12]Chang Fei,Zhang Lin-hong,Xu Wu-ping,et al.microRNA-9attenuates amyloid beta-induced synaptotoxicity by targeting Calcium/calmodulin-dependent protein kinase kinase 2[J].Molecular Medicine Reports,2014,9(5):1917-1922.
[13]Kong Y,Wu J,Yuan L.MicroRNA expression analysis of adult-onset Drosophila Alzheimer's disease model[J].Current Alzheimer Research,2014,11(9):882-891.
[14]Cai L,Cai X.Up-regulation of miR-9 expression predicate advanced clinicopathological features and poor prognosis in patients with hepatocellular carcinoma[J].Diagn Pathol,2014,9(12):1000.
[15]Higashi T,Hayashi H,Ishimoto T,et al.miR-9-3p plays a tumour-suppressor role by targeting TAZ(WWTR1)in hepatocellular carcinoma cells[J].British Journal of Cancer,2015,113(2):252-258.
[16]Zhang Jiang-bo,Cheng Jin,Zeng Zhen-zhen,et al.Comprehensive profiling of novel microRNA-9 targets and a tumor suppressor role of microRNA-9 via targeting IGF2BP1 in hepatocellular carcinoma[J].Oncotarget,2015,6(39):42040-42052.
[17]Drakaki A,Hatziapostolou M,Polytarchou C,et al.Functional microRNA high throughput screening reveals miR-9 as a central regulator of liver oncogenesis by affecting the PPARA-CDH1 pathway[J].BMC Cancer,2015,15(7):542.
[18]Sun Zhao,Han Qin,Zhou Na,et al.MicroRNA-9 enhances migration and invasion through KLF17 in hepatocellular carcinoma[J].Molecular Oncology,2013,7(5):884-894.
[19]Liu Q-y,Chang M-n,Lei J-x,et al.Identification of microRNAs involved in Alzheimer's progression using a rabbit model of the disease[J].American Journal of Neurodegenerative Disease,2014,3(1):33-44.
[20]Romano GL,Platania CB,Forte S,et al.MicroRNA target prediction in glaucoma[J].Prog Brain Res,2015,220(6):217-240.
[21]Kropivsek K,Pickford J,Carter DA.Postnatal dynamics of Zeb2 expression in rat brain:analysis of novel 3'UTR sequence reveals a miR-9interacting site[J].Journal of Molecular Neuroscience,2014,52(1):138-147.
[22]Sim SE,Lim CS,Kim JI,et al.The Brain-Enriched MicroRNAmiR-9-3p regulates synaptic plasticity and memory[J].Journal of Neuroscience,2016,36(33):8641-8652.
[23]Liu Zhen,Wang Cun-fu,Wang Xiao,et al.Therapeutic effects of transplantation of As-MiR-937-Expressing mesenchymal stem cells in murine model of alzheimer's disease[J].Cellular Physiology and Biochemistry,2015,37(1):321-330.
[24]Jung HJ,Coffinier C,Choe Y,et al.Regulation of prelamin Abut not lamin C by miR-9,a brain-specific microRNA[J].Proceedings of the National Academy of Sciences of the United States of America,2012,109(7):E423-E431.