早老素-1 p.Met139Ile突变对SH-SY5Y细胞模型相关蛋白与miR-34a表达影响  

INFLUENCE OF PRESENILIN-1 P.MET139ILE MUTATION ON THE EXPRESSION OF RELATED PROTEINS AND MIR-34A IN SH-SY5Y CELL MODEL

作　　者：孙雪纯 杨绍楠 王源[1] 张雅梦 潘旭东[1] 马爱军[1] SUN Xuechun;YANG Shaonan;WANG Yuan;ZHANG Yameng;PAN Xudong;MA Aijun(Department of Neurology,The Affiliated Hospital of Qingdao University,Qingdao 266100,China)

机构地区：[1]青岛大学附属医院神经内科,山东青岛266100

出　　处：《青岛大学学报（医学版）》2021年第1期87-90,共4页Journal of Qingdao University(Medical Sciences)

基　　金：山东省科技发展计划项目(2012G0021842)。

摘　　要：目的探讨早老素-1(PSEN-1)p.Met139Ile突变对早发性阿尔茨海默病(EOAD)人神经母细胞瘤细胞(SH-SY5Y细胞)模型中相关蛋白及microRNA-34a(miR-34a)表达的影响。方法分别将携带有阴性对照空载体的慢病毒(NC组)、野生型PSEN-1的慢病毒(WT组)以及突变型PSEN-1 p.Met139Ile(c.417G>C)的慢病毒(MT组)转染至SH-SY5Y细胞,以未感染慢病毒细胞作为正常组(N组)。ELISA方法检测各组细胞β-淀粉样蛋白表达(Aβ42/Aβ40值),Western blot方法检测各组PSEN-1、淀粉样蛋白前体(APP)、NOTCH-1蛋白的表达,qRT-PCR方法检测各组miR-34a的表达。结果与N组、NC组、WT组比较,MT组Aβ42/Aβ40值、PSEN-1蛋白、APP蛋白及miR-34a表达升高,而NOTCH-1蛋白表达下调,差异有显著性(F=28.540~425.600,q=7.349~30.180,P<0.05),N组、NC组、WT组各指标比较差异无显著性(P>0.05)。结论PSEN-1 p.Met139Ile为致病性突变位点,可引起SH-SY5Y细胞模型中相关蛋白及miR-34a的异常表达。Objective To investigate the influence of presenilin-1(PSEN-1)p.Met139Ile mutation on the expression of related proteins and microRNA-34a(miR-34a)in a model of human neuroblastoma SH-SY5Y cells in early-onset Alzheimer’s di-sease.Methods SH-SY5Y cells were transfected with the lentivirus carrying negative control(NC group),the lentivirus carrying wild-type PSEN-1(WT group),and the lentivirus carrying mutant-type PSEN-1 p.Met139Ile(c.417G>C)(MT group),and the cells not transfected with lentivirus were established as normal group(N group).ELISA was used to measure the expression of amyloidβ-proteins(Aβ42/Aβ40 ratio);Western blot was used to measure the protein expression of PSEN-1,amyloid precursor protein(APP),and NOTCH-1;qRT-PCR was used to measure the expression of miR-34a.Results Compared with the N,NC,and WT groups,the MT group had significant increases in Aβ42/Aβ40 ratio and expression of PSEN-1,APP,and miR-34a and a significant reduction in the expression of NOTCH-1(F=28.540-425.600,q=7.349-30.180,P<0.05),while there were no significant differences in these indices between the N,NC,and WT groups(P>0.05).Conclusion PSEN-1 p.Met139Ile is a pathogenic mutation site and can cause abnormal expression of related proteins and miR-34a in the model of SH-SY5Y cells.

关 键 词：早老素-1 阿尔茨海默病 淀粉样β肽类 微RNAS 

分 类 号：R745.7[医药卫生—神经病学与精神病学]