miR-15b-5p targeting amyloid precursor protein is involved in the anti-amyloid eflect of curcumin in swAPP695-HEK293 cells
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摘要
Curcumin exerts a neuroprotective effect on Alzheimer's disease; however, it is not known whether microRNAs are involved in this protective effect. This study was conducted using swAPP695-HEK293 cells as an Alzheimer's disease cell model. swAPP695-HEK293 cells were treated with 0, 0.5, 1, 2, 5, and 10 μM curcumin for 24 hours. The changes in miR-15b-5p, miR-19a-3p, miR-195-5p, miR-101-3p, miR-216b-5p, miR-16-5p and miR-185-5p expression were assessed by real-time quantitative polymerase chain reaction. The mRNA and protein levels of amyloid precursor protein, amyloid-β40 and amyloid-β42 were evaluated by quantitative real-time polymerase chain reaction, western blot assays and enzyme-linked immunosorbent assays. swAPP695-HEK293 cells were transfected with miR-15b-5p mimic, or treated with 1 μM curcumin 24 hours before miR-15b-5p inhibitor transfection. The effects of curcumin on amyloid precursor protein, amyloid-β40 and amyloid-β42 levels were evaluated by western blot assays and enzyme-linked immunosorbent assay. Luciferase assays were used to analyze the interaction between miR-15b-5p and the 3′-untranslated region of amyloid precursor protein. The results show that amyloid precursor protein and amyloid-β expression were enhanced in swAPP695-HEK293 cells compared with HEK293parental cells. Curcumin suppressed the expression of amyloid precursor protein and amyloid-β and up-regulated the expression of miR-15b-5p in swAPP695-HEK293 cells. In addition, we found a negative association of miR-15b-5p expression with amyloid precursor protein and amyloid-β levels in the curcumin-treated cells. Luciferase assays revealed that miR-15b-5p impaired the luciferase activity of the plasmid harboring the 3′-untranslated region of amyloid precursor protein. These findings indicate that curcumin down-regulates the expression of amyloid precursor protein and amyloid-β in swAPP695-HEK293 cells, which was partially mediated by miR-15b-5p via targeting of the 3′-untranslated region of amyloid precursor protein.
Curcumin exerts a neuroprotective effect on Alzheimer's disease; however, it is not known whether microRNAs are involved in this protective effect. This study was conducted using swAPP695-HEK293 cells as an Alzheimer's disease cell model. swAPP695-HEK293 cells were treated with 0, 0.5, 1, 2, 5, and 10 μM curcumin for 24 hours. The changes in miR-15b-5p, miR-19a-3p, miR-195-5p, miR-101-3p, miR-216b-5p, miR-16-5p and miR-185-5p expression were assessed by real-time quantitative polymerase chain reaction. The mRNA and protein levels of amyloid precursor protein, amyloid-β40 and amyloid-β42 were evaluated by quantitative real-time polymerase chain reaction, western blot assays and enzyme-linked immunosorbent assays. swAPP695-HEK293 cells were transfected with miR-15b-5p mimic, or treated with 1 μM curcumin 24 hours before miR-15b-5p inhibitor transfection. The effects of curcumin on amyloid precursor protein, amyloid-β40 and amyloid-β42 levels were evaluated by western blot assays and enzyme-linked immunosorbent assay. Luciferase assays were used to analyze the interaction between miR-15b-5p and the 3′-untranslated region of amyloid precursor protein. The results show that amyloid precursor protein and amyloid-β expression were enhanced in swAPP695-HEK293 cells compared with HEK293parental cells. Curcumin suppressed the expression of amyloid precursor protein and amyloid-β and up-regulated the expression of miR-15b-5p in swAPP695-HEK293 cells. In addition, we found a negative association of miR-15b-5p expression with amyloid precursor protein and amyloid-β levels in the curcumin-treated cells. Luciferase assays revealed that miR-15b-5p impaired the luciferase activity of the plasmid harboring the 3′-untranslated region of amyloid precursor protein. These findings indicate that curcumin down-regulates the expression of amyloid precursor protein and amyloid-β in swAPP695-HEK293 cells, which was partially mediated by miR-15b-5p via targeting of the 3′-untranslated region of amyloid precursor protein.
Curcumin exerts a neuroprotective effect on Alzheimer's disease; however, it is not known whether microRNAs are involved in this protective effect. This study was conducted using swAPP695-HEK293 cells as an Alzheimer's disease cell model. swAPP695-HEK293 cells were treated with 0, 0.5, 1, 2, 5, and 10 μM curcumin for 24 hours. The changes in miR-15b-5p, miR-19a-3p, miR-195-5p, miR-101-3p, miR-216b-5p, miR-16-5p and miR-185-5p expression were assessed by real-time quantitative polymerase chain reaction. The mRNA and protein levels of amyloid precursor protein, amyloid-β40 and amyloid-β42 were evaluated by quantitative real-time polymerase chain reaction, western blot assays and enzyme-linked immunosorbent assays. swAPP695-HEK293 cells were transfected with miR-15b-5p mimic, or treated with 1 μM curcumin 24 hours before miR-15b-5p inhibitor transfection. The effects of curcumin on amyloid precursor protein, amyloid-β40 and amyloid-β42 levels were evaluated by western blot assays and enzyme-linked immunosorbent assay. Luciferase assays were used to analyze the interaction between miR-15b-5p and the 3′-untranslated region of amyloid precursor protein. The results show that amyloid precursor protein and amyloid-β expression were enhanced in swAPP695-HEK293 cells compared with HEK293parental cells. Curcumin suppressed the expression of amyloid precursor protein and amyloid-β and up-regulated the expression of miR-15b-5p in swAPP695-HEK293 cells. In addition, we found a negative association of miR-15b-5p expression with amyloid precursor protein and amyloid-β levels in the curcumin-treated cells. Luciferase assays revealed that miR-15b-5p impaired the luciferase activity of the plasmid harboring the 3′-untranslated region of amyloid precursor protein. These findings indicate that curcumin down-regulates the expression of amyloid precursor protein and amyloid-β in swAPP695-HEK293 cells, which was partially mediated by miR-15b-5p via targeting of the 3′-untranslated region of amyloid precursor protein.
引文
Alzheimer’s Association(2018)2018 Alzheimer’s disease facts and figures.Alzheimers Dement 14:367-429.
Austin SA,Sens MA,Combs CK(2009)Amyloid precursor protein mediates a tyrosine kinase-dependent activation response in endothelial cells.J Neurosci 29:14451-14462.
Bekris LM,Leverenz JB(2015)The biomarker and therapeutic potential of miRNA in Alzheimer’s disease.Neurodegener Dis Manag 5:61-74.
Boyanapalli SS,Kong AT(2015)“Curcumin,the King of Spices”:epigenetic regulatory mechanisms in the prevention of cancer,neurological,and inflammatory diseases.Curr Pharmacol Rep1:129-139.
Chang L,Yuan Y,Li C,Guo T,Qi H,Xiao Y,Dong X,Liu Z,Liu Q(2016)Upregulation of SNHG6 regulates ZEB1 expression by competitively binding miR-101-3p and interacting with UPF1 in hepatocellular carcinoma.Cancer Lett 383:183-194.
Chen M,Du ZY,Zheng X,Li DL,Zhou RP,Zhang K(2018)Use of?curcumin?in diagnosis,prevention,and treatment of Alzheimer’s disease.Neural Regen Res 13:742-752.
Cox SJ,Rodriguez Camargo DC,Lee YH,Reif B,Ivanova M,Ramamoorthy A(2018)Amyloid aggregation of hIAPP,Aβ,and calcitonin altered by a curcumin derivative.Biophys J 114:226a.
Galimberti D,Villa C,Fenoglio C,Serpente M,Ghezzi L,Cioffi SM,Arighi A,Fumagalli G,Scarpini E(2014)Circulating miRNAs as potential biomarkers in Alzheimer’s disease.J Alzheimers Dis42:1261-1267.
Goozee KG,Shah TM,Sohrabi HR,Rainey-Smith SR,Brown B,Verdile G,Martins RN(2016)Examining the potential clinical value of curcumin in the prevention and diagnosis of Alzheimer’s disease.Br J Nutr 115:449-465.
Jiang Y,Mullaney KA,Peterhoff CM,Che S,Schmidt SD,Boyer-Boiteau A,Ginsberg SD,Cataldo AM,Mathews PM,Nixon RA(2010)Alzheimer’s-related endosome dysfunction in Down syndrome is Abeta-independent but requires APP and is reversed by BACE-1 inhibition.Proc Natl Acad Sci U S A 107:1630-1635.
Jiao DM,Yan L,Wang LS,Hu HZ,Tang XL,Chen J,Wang J,Li Y,Chen QY(2017)Exploration of inhibitory mechanisms of curcumin in lung cancer metastasis using a miRNA-transcription factor-target gene network.PLoS One 12:e0172470.
Kumar P,Dezso Z,MacKenzie C,Oestreicher J,Agoulnik S,Byrne M,Bernier F,Yanagimachi M,Aoshima K,Oda Y(2013)Circulating miRNA biomarkers for Alzheimer’s disease.PLoS One8:e69807.
Lin W,Yang LK,Zhu J,Wang YH,Dong JR,Chen T,Wang D,Xu XM,Sun SB,Zhang L(2018)Deep brain stimulation for the treatment of moderate-to-severe Alzheimer’s disease:Study protocol for a prospective self-controlled trial.Clin Trials Degener Dis 3:66-70.
Liu CG,Wang JL,Li L,Wang PC(2014)MicroRNA-384 regulates both amyloid precursor protein and beta-secretase expression and is a potential biomarker for Alzheimer’s disease.Int J Mol Med 34:160-166.
Liu H,Li Z,Qiu D,Gu Q,Lei Q,Mao L(2010)The inhibitory effects of different curcuminoids on beta-amyloid protein,beta-amyloid precursor protein and beta-site amyloid precursor protein cleaving enzyme 1 in swAPP HEK293 cells.Neurosci Lett485:83-88.
Livak KJ,Schmittgen TD(2001)Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T))Method.Methods 25:402-408.
Long JM,Lahiri DK(2011)MicroRNA-101 downregulates Alzheimer’s amyloid-beta precursor protein levels in human cell cultures and is differentially expressed.Biochem Biophys Res Commun 404:889-895.
Long JM,Ray B,Lahiri DK(2012)MicroRNA-153 physiologically inhibits expression of amyloid-beta precursor protein in cultured human fetal brain cells and is dysregulated in a subset of Alzheimer disease patients.J Biol Chem 287:31298-31310.
Lugli G,Cohen AM,Bennett DA,Shah RC,Fields CJ,Hernandez AG,Smalheiser NR(2015)Plasma exosomal miRNAs in persons with and without Alzheimer disease:altered expression and prospects for biomarkers.PLoS One 10:e0139233.
Lukiw WJ,Alexandrov PN(2012)Regulation of complement factor H(CFH)by multiple miRNAs in Alzheimer’s disease(AD)brain.Mol Neurobiol 46:11-19.
Ma J,Fang B,Zeng F,Pang H,Zhang J,Shi Y,Wu X,Cheng L,Ma C,Xia J,Wang Z(2014)Curcumin inhibits cell growth and invasion through up-regulation of miR-7 in pancreatic cancer cells.Toxicol Lett 231:82-91.
Momtazi AA,Derosa G,Maffioli P,Banach M,Sahebkar A(2016)Role of microRNAs in the therapeutic effects of curcumin in non-cancer diseases.Mol Diagn Ther 20:335-345.
Nhan HS,Chiang K,Koo EH(2015)The multifaceted nature of amyloid precursor protein and its proteolytic fragments:friends and foes.Acta Neuropathol 129:1-19.
Patel N,Hoang D,Miller N,Ansaloni S,Huang Q,Rogers JT,Lee JC,Saunders AJ(2008)MicroRNAs can regulate human APPlevels.Mol Neurodegener 3:10.
Rockenstein E,Mallory M,Mante M,Sisk A,Masliaha E(2001)Early formation of mature amyloid-beta protein deposits in a mutant APP transgenic model depends on levels of Abeta(1-42).J Neurosci Res 66:573-582.
Rupaimoole R,Slack FJ(2017)MicroRNA therapeutics:towards a new era for the management of cancer and other diseases.Nat Rev Drug Discov 16:203-222.
Sandur SK,Pandey MK,Sung B,Ahn KS,Murakami A,Sethi G,Limtrakul P,Badmaev V,Aggarwal BB(2007)Curcumin,demethoxycurcumin,bisdemethoxycurcumin,tetrahydrocurcumin and turmerones differentially regulate anti-inflammatory and anti-proliferative responses through a ROS-independent mechanism.Carcinogenesis 28:1765-1773.
Somparn P,Phisalaphong C,Nakornchai S,Unchern S,Morales NP(2007)Comparative antioxidant activities of curcumin and its demethoxy and hydrogenated derivatives.Biol Pharm Bull 30:74-78.
Sun CC,Zhang L,Li G,Li SJ,Chen ZL,Fu YF,Gong FY,Bai T,Zhang DY,Wu QM,Li DJ(2017)The lncRNA PDIA3P interacts with miR-185-5p to modulate oral squamous cell carcinoma progression by targeting Cyclin D2.Mol Ther Nucleic Acids 9:100-110.
Toden S,Okugawa Y,Buhrmann C,Nattamai D,Anguiano E,Baldwin N,Shakibaei M,Boland CR,Goel A(2015)Novel evidence for curcumin and boswellic acid-induced chemoprevention through regulation of miR-34a and miR-27a in colorectal cancer.Cancer Prev Res(Phila)8:431-443.
Wan Y,Liu Y,Wang X,Wu J,Liu K,Zhou J,Liu L,Zhang C(2015)Identification of differential microRNAs in cerebrospinal fluid and serum of patients with major depressive disorder.PLoS One10:e0121975.
Wang XH,Wang TL(2018)MicroRNAs of microglia:Wrestling with central nervous system disease.Neural Regen Res 13:2067-2072.
Wu Y,Deng Y,Zhang S,Luo Y,Cai F,Zhang Z,Zhou W,Li T,Song W(2015)Amyloid-beta precursor protein facilitates the regulator of calcineurin 1-mediated apoptosis by downregulating proteasome subunit alpha type-5 and proteasome subunit beta type-7.Neurobiol Aging 36:169-177.
Xiao Q,Yan P,Ma X,Liu H,Perez R,Zhu A,Gonzales E,Tripoli DL,Czerniewski L,Ballabio A,Cirrito JR,Diwan A,Lee J-M(2015)Neuronal-targeted TFEB accelerates lysosomal degradation of APP,reducing Aβgeneration and amyloid plaque pathogenesis.J Neurosci 35:12137-12151.
Ye M,Zhang J,Zhang J,Miao Q,Yao L,Zhang J(2015)Curcumin promotes apoptosis by activating the p53-miR-192-5p/215-XIAPpathway in non-small cell lung cancer.Cancer Lett 357:196-205.
Zhang J,Song Y,Zhang C,Zhi X,Fu H,Ma Y,Chen Y,Pan F,Wang K,Ni J,Jin W,He X,Su H,Cui D(2015)Circulating MiR-16-5p and MiR-19b-3p as two novel potential biomarkers to indicate progression of gastric cancer.Theranostics 5:733-745.
Austin SA,Sens MA,Combs CK(2009)Amyloid precursor protein mediates a tyrosine kinase-dependent activation response in endothelial cells.J Neurosci 29:14451-14462.
Bekris LM,Leverenz JB(2015)The biomarker and therapeutic potential of miRNA in Alzheimer’s disease.Neurodegener Dis Manag 5:61-74.
Boyanapalli SS,Kong AT(2015)“Curcumin,the King of Spices”:epigenetic regulatory mechanisms in the prevention of cancer,neurological,and inflammatory diseases.Curr Pharmacol Rep1:129-139.
Chang L,Yuan Y,Li C,Guo T,Qi H,Xiao Y,Dong X,Liu Z,Liu Q(2016)Upregulation of SNHG6 regulates ZEB1 expression by competitively binding miR-101-3p and interacting with UPF1 in hepatocellular carcinoma.Cancer Lett 383:183-194.
Chen M,Du ZY,Zheng X,Li DL,Zhou RP,Zhang K(2018)Use of?curcumin?in diagnosis,prevention,and treatment of Alzheimer’s disease.Neural Regen Res 13:742-752.
Cox SJ,Rodriguez Camargo DC,Lee YH,Reif B,Ivanova M,Ramamoorthy A(2018)Amyloid aggregation of hIAPP,Aβ,and calcitonin altered by a curcumin derivative.Biophys J 114:226a.
Galimberti D,Villa C,Fenoglio C,Serpente M,Ghezzi L,Cioffi SM,Arighi A,Fumagalli G,Scarpini E(2014)Circulating miRNAs as potential biomarkers in Alzheimer’s disease.J Alzheimers Dis42:1261-1267.
Goozee KG,Shah TM,Sohrabi HR,Rainey-Smith SR,Brown B,Verdile G,Martins RN(2016)Examining the potential clinical value of curcumin in the prevention and diagnosis of Alzheimer’s disease.Br J Nutr 115:449-465.
Jiang Y,Mullaney KA,Peterhoff CM,Che S,Schmidt SD,Boyer-Boiteau A,Ginsberg SD,Cataldo AM,Mathews PM,Nixon RA(2010)Alzheimer’s-related endosome dysfunction in Down syndrome is Abeta-independent but requires APP and is reversed by BACE-1 inhibition.Proc Natl Acad Sci U S A 107:1630-1635.
Jiao DM,Yan L,Wang LS,Hu HZ,Tang XL,Chen J,Wang J,Li Y,Chen QY(2017)Exploration of inhibitory mechanisms of curcumin in lung cancer metastasis using a miRNA-transcription factor-target gene network.PLoS One 12:e0172470.
Kumar P,Dezso Z,MacKenzie C,Oestreicher J,Agoulnik S,Byrne M,Bernier F,Yanagimachi M,Aoshima K,Oda Y(2013)Circulating miRNA biomarkers for Alzheimer’s disease.PLoS One8:e69807.
Lin W,Yang LK,Zhu J,Wang YH,Dong JR,Chen T,Wang D,Xu XM,Sun SB,Zhang L(2018)Deep brain stimulation for the treatment of moderate-to-severe Alzheimer’s disease:Study protocol for a prospective self-controlled trial.Clin Trials Degener Dis 3:66-70.
Liu CG,Wang JL,Li L,Wang PC(2014)MicroRNA-384 regulates both amyloid precursor protein and beta-secretase expression and is a potential biomarker for Alzheimer’s disease.Int J Mol Med 34:160-166.
Liu H,Li Z,Qiu D,Gu Q,Lei Q,Mao L(2010)The inhibitory effects of different curcuminoids on beta-amyloid protein,beta-amyloid precursor protein and beta-site amyloid precursor protein cleaving enzyme 1 in swAPP HEK293 cells.Neurosci Lett485:83-88.
Livak KJ,Schmittgen TD(2001)Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T))Method.Methods 25:402-408.
Long JM,Lahiri DK(2011)MicroRNA-101 downregulates Alzheimer’s amyloid-beta precursor protein levels in human cell cultures and is differentially expressed.Biochem Biophys Res Commun 404:889-895.
Long JM,Ray B,Lahiri DK(2012)MicroRNA-153 physiologically inhibits expression of amyloid-beta precursor protein in cultured human fetal brain cells and is dysregulated in a subset of Alzheimer disease patients.J Biol Chem 287:31298-31310.
Lugli G,Cohen AM,Bennett DA,Shah RC,Fields CJ,Hernandez AG,Smalheiser NR(2015)Plasma exosomal miRNAs in persons with and without Alzheimer disease:altered expression and prospects for biomarkers.PLoS One 10:e0139233.
Lukiw WJ,Alexandrov PN(2012)Regulation of complement factor H(CFH)by multiple miRNAs in Alzheimer’s disease(AD)brain.Mol Neurobiol 46:11-19.
Ma J,Fang B,Zeng F,Pang H,Zhang J,Shi Y,Wu X,Cheng L,Ma C,Xia J,Wang Z(2014)Curcumin inhibits cell growth and invasion through up-regulation of miR-7 in pancreatic cancer cells.Toxicol Lett 231:82-91.
Momtazi AA,Derosa G,Maffioli P,Banach M,Sahebkar A(2016)Role of microRNAs in the therapeutic effects of curcumin in non-cancer diseases.Mol Diagn Ther 20:335-345.
Nhan HS,Chiang K,Koo EH(2015)The multifaceted nature of amyloid precursor protein and its proteolytic fragments:friends and foes.Acta Neuropathol 129:1-19.
Patel N,Hoang D,Miller N,Ansaloni S,Huang Q,Rogers JT,Lee JC,Saunders AJ(2008)MicroRNAs can regulate human APPlevels.Mol Neurodegener 3:10.
Rockenstein E,Mallory M,Mante M,Sisk A,Masliaha E(2001)Early formation of mature amyloid-beta protein deposits in a mutant APP transgenic model depends on levels of Abeta(1-42).J Neurosci Res 66:573-582.
Rupaimoole R,Slack FJ(2017)MicroRNA therapeutics:towards a new era for the management of cancer and other diseases.Nat Rev Drug Discov 16:203-222.
Sandur SK,Pandey MK,Sung B,Ahn KS,Murakami A,Sethi G,Limtrakul P,Badmaev V,Aggarwal BB(2007)Curcumin,demethoxycurcumin,bisdemethoxycurcumin,tetrahydrocurcumin and turmerones differentially regulate anti-inflammatory and anti-proliferative responses through a ROS-independent mechanism.Carcinogenesis 28:1765-1773.
Somparn P,Phisalaphong C,Nakornchai S,Unchern S,Morales NP(2007)Comparative antioxidant activities of curcumin and its demethoxy and hydrogenated derivatives.Biol Pharm Bull 30:74-78.
Sun CC,Zhang L,Li G,Li SJ,Chen ZL,Fu YF,Gong FY,Bai T,Zhang DY,Wu QM,Li DJ(2017)The lncRNA PDIA3P interacts with miR-185-5p to modulate oral squamous cell carcinoma progression by targeting Cyclin D2.Mol Ther Nucleic Acids 9:100-110.
Toden S,Okugawa Y,Buhrmann C,Nattamai D,Anguiano E,Baldwin N,Shakibaei M,Boland CR,Goel A(2015)Novel evidence for curcumin and boswellic acid-induced chemoprevention through regulation of miR-34a and miR-27a in colorectal cancer.Cancer Prev Res(Phila)8:431-443.
Wan Y,Liu Y,Wang X,Wu J,Liu K,Zhou J,Liu L,Zhang C(2015)Identification of differential microRNAs in cerebrospinal fluid and serum of patients with major depressive disorder.PLoS One10:e0121975.
Wang XH,Wang TL(2018)MicroRNAs of microglia:Wrestling with central nervous system disease.Neural Regen Res 13:2067-2072.
Wu Y,Deng Y,Zhang S,Luo Y,Cai F,Zhang Z,Zhou W,Li T,Song W(2015)Amyloid-beta precursor protein facilitates the regulator of calcineurin 1-mediated apoptosis by downregulating proteasome subunit alpha type-5 and proteasome subunit beta type-7.Neurobiol Aging 36:169-177.
Xiao Q,Yan P,Ma X,Liu H,Perez R,Zhu A,Gonzales E,Tripoli DL,Czerniewski L,Ballabio A,Cirrito JR,Diwan A,Lee J-M(2015)Neuronal-targeted TFEB accelerates lysosomal degradation of APP,reducing Aβgeneration and amyloid plaque pathogenesis.J Neurosci 35:12137-12151.
Ye M,Zhang J,Zhang J,Miao Q,Yao L,Zhang J(2015)Curcumin promotes apoptosis by activating the p53-miR-192-5p/215-XIAPpathway in non-small cell lung cancer.Cancer Lett 357:196-205.
Zhang J,Song Y,Zhang C,Zhi X,Fu H,Ma Y,Chen Y,Pan F,Wang K,Ni J,Jin W,He X,Su H,Cui D(2015)Circulating MiR-16-5p and MiR-19b-3p as two novel potential biomarkers to indicate progression of gastric cancer.Theranostics 5:733-745.