摘要
目的探讨长链非编码RNA(lncRNA)Sox2OT过表达对Aβ1-42诱导PC12细胞损伤及PI3K/Akt通路的影响。方法选取大鼠嗜铬神经瘤细胞株(PC12),利用Aβ1-42对PC12细胞进行处理,建立AD的细胞模型。将Aβ1-42诱导前的PC12细胞设为空白组;将Aβ1-42诱导后的PC12细胞分为control组、Sox2OT过表达(p-Sox2OT)组、p-Sox2OT空载体(p-NC)组、抑制Sox2OT表达(si-Sox2OT)组和si-Sox2OT空载体(si-NC)组。使用噻唑蓝(MTT)方法对细胞的增殖活性进行检测;采用流式细胞术对转染后的细胞周期和凋亡率进行检测;采用Western blot检测PI3K/Akt通路蛋白的表达。结果MTT结果显示,与空白组(99.67±10.50)相比,control组(29.33±5.51)的细胞增殖率显著降低(t=10.27,P<0.05)。RT-qPCR结果显示,与control组(0.52±0.06)相比,p-Sox2OT组(2.19±0.16)中的Sox2OT的mRNA表达水平显著升高(t=16.93,P<0.05),si-Sox2OT组(0.22±0.02)中的Sox2OT的mRNA表达水平显著下降(t=15.28,P<0.05);与p-NC组(0.53±0.12)相比,p-Sox2OT组(2.19±0.16)中的Sox2OT的mRNA表达水平显著升高(t=16.25,P<0.05);与si-NC组(0.51±0.09)相比,si-Sox2OT组(0.22±0.02)中的Sox2OT的mRNA表达水平显著下降(t=16.93,P<0.05);control组与p-NC组以及si-NC组间的差异无统计学意义(P>0.05)。此外,p-Sox2OT组细胞的增殖能力(145.00±5.12)显著高于si-Sox2OT组(23.33±4.93)、control组(55.00±5.00)、si-NC组(57.33±8.51)以及p-NC组(56.00±5.57)(t=29.65,21.78,27.55,21.35,均P<0.05)。Control组与p-NC组以及si-NC组间细胞增殖率的差异不具有统计学意义(P>0.05)。细胞周期检测实验显示,p-Sox2OT组G1期的细胞数量显著低于control组和p-NC(t=9.80,8.57;均P<0.05),而p-Sox2OT组G2期的细胞数量与control组和p-NC组相比却显著升高(t=11.02,10.25;均P<0.05);si-Sox2OT组G1期的细胞数量显著高于control组和si-NC组(t=8.22,3.11,均P<0.05),而G2期的细胞数量与control组和si-NC组相比却显著下降(t=6.32,5.33;均P<0.05);control组与p-NC组以及si-NC组间的细胞周期�
Objective To investigate the mechanism of lncRNA Sox2OT in patients with Alzheimer's disease(AD)induced by A type ofβpeptide(Aβ1-42).Methods Rat pheochromocytoma cells(PC12 cells)were selected and treated by Aβ1-42 to establish PC12 cell model.PC12 cells were set as blank group before induction to verify the successful construction of the cell model.The induced PC12 cells were divided into control group,Sox2OT overexpression(p-Sox2OT)group,p-Sox2OT empty vector(p-NC)group,inhibited Sox2OT expression(si-Sox2OT)group and si-Sox2OT empty vector(si-NC)group.The proliferation activity of thiazole blue(MTT)was detected.Flow cytometry was used to detect the cell cycle and apoptosis rate after transfection.Results MTT results showed that compared with the blank group(99.67±10.50),the cell proliferation rate of the control group(29.33±5.51)was significantly reduced(t=10.27,P<0.05).RT-qPCR results showed that compared with the control group(0.52±0.06),the Sox2OT mRNA expression level in the p-Sox2OT group(2.19±0.16)was significantly increased(t=16.93,P<0.05).The mRNA expression level of Sox2OT in the si-Sox2OT group(0.22±0.02)decreased significantly(t=15.28,P<0.05).Compared with the p-NC group(0.53±0.12),The mRNA expression level of Sox2OT in the p-Sox2OT group(2.19±0.16)was significantly increased(t=16.25,P<0.05).Compared with the si-NC group(0.51±0.09),the mRNA expression level of Sox2OT in the si-Sox2OT group(0.22±0.02)was significantly decreased(t=16.93,P<0.05).The difference between the control group,the p-NC group and the si-NC group was not statistically significant(P>0.05).In addition,the cell proliferation ability of the p-Sox2OT group(145.00±5.12)was significantly higher than that of the si-Sox2OT group(23.33±4.93),control group(55.00±5.00),si-NC group(57.33±8.51)and p-NC group(56.00±5.57)(t=29.65,21.78,27.55,21.35,all P<0.05).The difference in cell proliferation rate between Control group,p-NC group and si-NC group was not statistically significant(P>0.05).Cell cycle detection experiments showed
作者
陈鹏
李龙
刘春华
张昭强
孙宪昌
Chen Peng;Li Long;Liu Chunhua;Zhang Zhaoqiang;Sun Xianchang(Department of Physiology,College of Basic Medicine,Shandong First Medical University&Shandong Academy of Medical Sciences,Taian 271016,China;Department of Oncology,Taian Tumour Prevention and Treatment Hospital,Taian 271016,China)
出处
《中华行为医学与脑科学杂志》
CAS
CSCD
北大核心
2020年第9期785-791,共7页
Chinese Journal of Behavioral Medicine and Brain Science
基金
山东省自然科学基金项目(ZR2019MH090)
山东省卫生与计划生育委员会项目(2015WS0107)。
