右美托咪定抑制DEAD/H盒解旋酶11反义核糖核酸1表达调控胶质瘤细胞增殖和凋亡  

Dexmedetomidine regulates proliferation and apoptosis of glioma cells by inhibiting DEAD/H box helicase 11 antisense RNA 1 expression

作　　者：张海盛[1] 姚泽宇[1] 张利亮 马建梅 孙永[2] Zhang Haisheng;Yao Zeyu;Zhang Liliang;Ma Jianmei;Sun Yong(Department of Anesthesiology,Qinghai Red Cross Hospital,Xining 810000,China;Department of Neurosurgery,Qinghai Red Cross Hospital,Xining 810000,China)

机构地区：[1]青海红十字医院麻醉科,西宁810000 [2]青海红十字医院神经外科,西宁810000

出　　处：《中华实验外科杂志》2020年第7期1259-1262,共4页Chinese Journal of Experimental Surgery

基　　金：青海省医药卫生科技项目(2016-wjzd-ll)。

摘　　要：目的探讨右美托咪定对SHG-44胶质瘤细胞增殖和凋亡的影响及其机制。方法采用0.1 μmol/L(低浓度组)、1.0 μmol/L(中浓度组)和10.0 μmol/L(高浓度组)右美托咪定处理SHG-44胶质瘤细胞(购自中国科学院细胞库)。将si-DEAD/H盒解旋酶11反义核糖核酸1(DDX11-AS1)和pcDNA-DDX11-AS1分别转染至SHG-44胶质瘤细胞并采用10.0 μmol/L右美托咪定处理。采用噻唑蓝(MTT)法、克隆形成实验、流式细胞仪、实时荧光定量聚合酶链反应(RT-qPCR)和蛋白质印迹法(Western blot)检测各组细胞增殖、克隆形成数、凋亡率、DDX11-AS1、裂解的半胱氨酰天冬氨酸特异性蛋白酶-3(cleaved-Caspase-3)、Caspase-3前体(pro-Caspase-3)表达。采用t检验或单因素方差分析。结果低浓度组、中浓度组和高浓度组SHG-44胶质瘤细胞吸光度(A)490值、克隆形成数、DDX11-AS1和pro-Caspase-3表达依次降低[A490值为1.17±0.05、0.90±0.04、0.55±0.03,克隆形成数为(108.67±3.09)、(82.00±2.45)、(55.00±2.16)个,DDX11-AS1为0.96±0.04、0.68±0.03、0.36±0.02,pro-Caspase-3为0.72±0.04、0.48±0.02、0.24±0.02],凋亡率和cleaved-Caspase-3表达依次升高[凋亡率为(6.38±0.18)%、(16.78±0.71)%、(21.77±0.77)%,cleaved-Caspase-3为0.23±0.01、0.45±0.02、0.64±0.03],差异有统计学意义(F=140.693、198.187、262.395、321.333、611.199、135.429,P<0.05)。si-DDX11-AS1组SHG-44胶质瘤细胞A490值、克隆形成数和pro-Caspase-3表达低于si-NC组SHG-44胶质瘤细胞[0.72±0.03比1.20±0.05、(65.00±2.45)个比(110.33±4.78)个、0.34±0.02比0.73±0.04,t=14.258、14.617、15.105,P<0.05],凋亡率和cleaved-Caspase-3高于si-NC组SHG-44胶质瘤细胞[(18.38±0.53)%比(6.45±0.20)%、0.53±0.02比0.21±0.01,t=36.477、24.787,P<0.05]。高浓度右美托咪定+pcDNA-DDX11-AS1组SHG-44胶质瘤细胞A490值、克隆形成数和pro-Caspase-3表达高于高浓度右美托咪定+pcDNA组[0.98±0.04比0.54±0.03、(93.33±3.09)个比(56.00±2.16)个、0.63±0.03比0.24±0.01,tObjective:To study the effect of dexmedetomidine on the proliferation and apoptosis of glioma cells and its mechanism.Methods:Low(0.1μmol/L),medium(1.0μmol/L)and high(10.0μmol/L)concentrations of dexmedetomidine were used to treat SHG-44 glioma cells.Si-DEAD/H box helicase 11 antisense RNA 1(DDX11-AS1)and pcDNA-DDX11-AS1 were respectively transfected into SHG-44 glioma cells and treated with 10.0μmol/L dexmedetomidine.The tetramethylazolium salt method,clone formation experiment,flow cytometry,real-time fluorescent quantitative polymerase chain reaction and Western blotting were used to detect cell proliferation and clone formation,apoptosis rate,DDX11-AS1,cleaved-cysteinyl aspartate-specific protease(Caspase)-3,pro-Caspase-3 expression.The t test or one-way analysis of variance was used to compare the measurement data differences.Results:The A490 value,the number of clones,the expression of DDX11-AS1 and pro-Caspase-3 in low concentration group,medium concentration group and high concentration group decreased sequentially(A490 values:1.17±0.05,0.90±0.04,0.55±0.03;the number of clone formation:108.67±3.09,82.00±2.45,55.00±2.16;DDX11-AS1:0.96±0.04,0.68±0.03,0.36±0.02;pro-Caspase-3:0.72±0.04,0.48±0.02,0.24±0.02),and the apoptotic rate and cleaved-Caspase-3 expression increased sequentially[apoptotic rate:(6.38±0.18)%,(16.78±0.71)%,(21.77±0.77)%;cleaved-Caspase-3:0.23±0.01,0.45±0.02,0.64±0.03].The A490 value,number of clone formation and pro-Caspase-3 expression in the si-DDX11-AS1 group were significantly lower than those in the si-NC group(0.72±0.03 vs.1.20±0.05,65.00±2.45 vs.110.33±4.78,0.34±0.02 vs.0.73±0.04,t=14.258,14.617,15.105,P<0.05),and apoptosis rate and cleaved-Caspase-3 were significantly higher[(18.38±0.53)%vs.(6.45±0.20)%,0.53±0.02 vs.0.21±0.01,t=36.477,24.787,P<0.05].The A490 value,number of clone formation and pro-Caspase-3 expression in the high concentration dexmedetomidine+pcDNA-DDX11-AS1 group were significantly higher than those in the high concentration dexmedet

关 键 词：胶质瘤 右美托咪定 DEAD/H盒解旋酶11反义核糖核酸1 增殖 脱噬作用 

分 类 号：R739.41[医药卫生—肿瘤] R614[医药卫生—临床医学]