TUG1对地塞米松诱导小鼠MC3T3-E1细胞凋亡的影响  

Effect of TUG1 on dexamethasone-induced apoptosis in mouse MC3T3-E1 cells

作　　者：刘亚楠 王娜[2] 梁秋华[2] 张艳芳 黄园园 于世鹏[2] LIU Yanan;WANG Na;LIANG Qiuhua;ZHANG Yanfang;HUANG Yuanyuan;YU Shipeng(School of Clinical Medicine,Jining Medical University,Jining 272013,China;Affiliated Hospital of Jining Medical University,Jining 272029,China)

机构地区：[1]济宁医学院临床医学院,济宁272013 [2]济宁医学院附属医院,济宁272029

出　　处：《济宁医学院学报》2019年第3期153-157,共5页Journal of Jining Medical University

基　　金：山东省医药卫生科技发展计划项目(2016WS0168)

摘　　要：目的探讨长链非编码RNA-TUG1对地塞米松诱导小鼠MC3T3-E1细胞凋亡的影响。方法体外培养MC3T3-E1细胞,建立细胞凋亡模型,诱导凋亡24h后qPCR检测TUG1的表达。将重组质粒导入MC3T3-E1细胞,获得稳定转染的细胞株,实验分为对照组、地塞米松组、地塞米松+TUG1过表达质粒组、地塞米松+阴性质粒组,qPCR检测TUG1的表达,流式细胞术检测细胞凋亡情况,免疫荧光观察Bcl-2的表达,计算平均荧光强度。结果细胞凋亡模型成功建立,荧光显微镜显示质粒转染MC3T3-E1细胞,qPCR验证质粒转染成功(P<0.05);地塞米松+TUG1过表达质粒组细胞凋亡明显低于地塞米松组和地塞米松+阴性质粒组,Bcl-2的表达在地塞米松+TUG1过表达质粒组明显高于地塞米松组和地塞米松+阴性质粒组,差异具有统计学意义(P<0.05)。结论TUG1可以通过上调Bcl-2的表达抑制地塞米松诱导的小鼠MC3T3-E1细胞凋亡,TUG1可能是糖皮质激素诱导的成骨细胞凋亡的保护性因素。Objective To investigate the effect of long non-coding RNA-TUG1 in dexamethasone-induced mice MC3T3-E1 cell apoptosis.Methods MC3T3-E1 cells were cultured in vitro to establish a model of apoptosis.After induction of apoptosis for 24h,qPCR was used to detect the expression of TUG1.The recombinant plasmid was introduced into MC3T3-E1 cells to obtain a stably transfected cell line.The experiment was divided into control group,dexamethasone group,dexamethasone+TUG1 overexpression plasmid group and dexamethasone+negative plasmid group.qPCR was used to detect the expression of TUG1,and the apoptosis was detected by flow cytometry.Bcl-2 expression was observed byimmunofluorescence,and the mean fluorescence intensity was calculated.Results The apoptosis model was successfully established.Fluorescence microscopy showed that plasmid transfected cells were correctly transfected with qPCR(P<0.05).Apoptosis of dexamethasone+TUG1 overexpressing plasmid group was significantly lower than that of dexamethasone group and dexamethasone+negative plasmid group.Meanwhile,the expression of Bcl-2 was significantly higher in the dexamethasone+TUG1 overexpression plasmid group than in the dexamethasone group and the dexamethasone+negative plasmid group(P<0.05).Conclusion TUG1 can inhibit the apoptosis of mouse MC3T3-E1 cells induced by dexamethasone by up-regulating the expression of Bcl-2.TUG1 may be a protective factor for glucocorticoid-induced osteoblast apoptosis.

关 键 词：成骨细胞 细胞凋亡 糖皮质激素 长链非编码RNA TUG1 

分 类 号：R34[医药卫生—基础医学]