长基因间非蛋白质编码核糖核酸01638对胶质瘤细胞增殖和凋亡影响及其机制  

Effects of long intergenic non-protein-coding RNA 01638 on proliferation and apoptosis of glioma cells and underlying mechanism

作　　者：李立超[1] 郑茂华[1] 谢民[1] 武志 雒以诚[1] 丁涛 Li Lichao;Zheng Maohua;Xie Min;Wu Zhi;Luo Yicheng;Ding Tao(Department of Neurosurgery,the First Hospital of Lanzhou University,Lanzhou 730030,China)

机构地区：[1]兰州大学第一医院神经外科,730030

出　　处：《中华实验外科杂志》2022年第9期1651-1654,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探讨长基因间非蛋白质编码核糖核酸01638(LINC01638)调控胶质瘤细胞增殖和凋亡机制。方法U251胶质瘤细胞购自中国科学院细胞库。选择2017年1月至2019年1月兰州大学第一医院诊治的颅脑胶质瘤患者及颅脑血肿清除术患者为研究对象。将U251细胞根据转染基因分为si-NC组、si-LINC01638组、pcDNA组、pcDNA-LINC01638组、miR-NC组、miR-647 mimics组、si-LINC01638+anti-miR-NC组和si-LINC01638+anti-miR-647组。实时荧光定量聚合酶链反应(RT-qPCR)检测LINC01638和微小核糖核酸647(miR-647)表达;细胞计数试剂盒-8(CCK-8)检测48 h细胞吸光度;流式细胞术检测细胞凋亡;蛋白质印迹法检测细胞周期蛋白D1(Cyclin D1)、细胞周期蛋白依赖性激酶抑制剂1A(p21)、B细胞淋巴瘤-2(bcl-2)及bcl-2相关X(bax)表达;双荧光素酶报告实验检测LINC01638和miR-647的靶向关系。两组间比较采用t检验;多组间比较采用单因素方差分析(两组间比较用SNK-q检验)。结果胶质瘤组织中LINC01638表达显著高于正常脑组织(2.96±0.27比0.95±0.08),miR-647表达显著低于正常脑组织(0.54±0.05比1.02±0.06),差异有统计学意义(t=31.921、27.485,P<0.05)。si-LINC01638组U251细胞LINC01638表达(0.49±0.03比0.97±0.07)、48 h细胞吸光度值(0.41±0.03比0.72±0.04)、Cyclin D1表达(0.24±0.02比0.69±0.04)显著低于si-NC组,p21表达(0.58±0.03比0.19±0.02)、细胞凋亡率[(22.63±3.18)%比(6.95±0.86)%]及bax表达(0.91±0.06比0.26±0.03)显著高于si-NC组,差异有统计学意义(t=14.238、18.432、20.399、16.981、19.322、24.876,P<0.05)。miR-647与WT-LINC01638共转染后U251细胞荧光素酶活性显著降低,转染pcDNA-LINC01638后miR-647低表达,转染si-LINC01638后miR-647高表达(P<0.05)。miR-647 mimics组U251细胞miR-647表达(2.71±0.08比1.01±0.03)、细胞凋亡率[(20.60±2.14)%比(6.12±0.39)%]、p21(0.51±0.06比0.18±0.02)和bax(0.83±0.07比0.23±0.04)表达显著高于miR-NC组,48 h细胞吸光度值(0.45±0.07�Objective To study the mechanism of long intergenic non-protein-coding RNA 01638 regulating glioma cell proliferation and apoptosis.Methods U251 glioma cells were purchased from the Cell Bank of the Chinese Academy of Sciences.Patients with craniocerebral glioma and those with craniocerebral hematoma from Jan.2017 to Jan.2019 were selected as research subjects.U251 cells were divided into si-NC group,si-LINC01638 group,pcDNA group,pcDNA-LINC01638 group,miR-NC group,miR-647 mimics group,si-LINC01638+anti-miR-NC group and si-LINC01638+anti-miR-647 group.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of LINC01638 and microribonucleic acid 647(miR-647).The cell counting kit-8(CCK-8)assay was used to detect the absorbance of cells at 48 h.The flow cytometry was used to detect cell apoptosis.Western blotting was used to detect the expression of Cyclin D1,cyclin dependent kinase inhibitor 1A(p21),B-cell lymphoma-2(bcl-2)and bcl-2-related X(bax).The dual-luciferase reporter assay was used to detect the targeting relationship between LINC01638 and miR-647.The t test was used for comparison between two groups,and the one-way analysis of variance was used for comparison between multiple groups(SNK-q test was used for comparison between two groups).Results The expression of LINC01638 in glioma tissue was significantly higher than that in normal brain tissue(2.96±0.27 vs.0.95±0.08),and that of miR-647 was significantly lower(0.54±0.05 vs.1.02±0.06)with statistical significances(t=31.921,27.485,P<0.05).The expression of LINC01638(0.49±0.03 vs.0.97±0.07),cell absorbance at 48 h(0.41±0.03 vs.0.72±0.04),and Cyclin D1 expression(0.24±0.02 vs.0.69±0.04)in U251 cells in si-LINC01638 group were significantly lower than those in si-NC group,while p21 expression(0.58±0.03 vs.0.19±0.02),apoptosis rate[(22.63±3.18)%vs.(6.95±0.86)%]and bax expression(0.91±0.06 vs.0.26±0.03)were significantly higher with statistical significances(t=14.238,18.432,20.399,16.981,19.322,24.876,P<0.05)

关 键 词：微小RNA 胶质瘤 增殖 凋亡 

分 类 号：R739.4[医药卫生—肿瘤]