沉默长链非编码RNA淋巴增强子结合因子1反义RNA1对人脐静脉内皮细胞的影响

Effect of silencing long non-coding RNA lymphoenhancer binding factor 1 antisense RNA1 on human umbilical vein endothelial cells

目的观察长链非编码RNA淋巴增强子结合因子1反义RNA1(LEF1-AS1)基因沉默后对人脐静脉内皮细胞(HUVEC)增殖、迁移和血管生成能力的影响。方法采用RNA干扰技术,在HUVEC(美国模式培养物集存库)中沉默LEF1-AS1基因的表达,细胞分为对照组与实验组,即NC组和si-LEF1-AS1组,实时荧光定量聚合酶链式反应(RT-qPCR)检测转染效果;细胞计数试剂盒(CCK-8)和Transwell分别检测细胞增殖和迁移力的变化;体外血管生成实验检测LEF1-AS1对血管生成能力的影响。两组间比较采用独立样本t检验。结果RT-qPCR结果显示,si-LEF1-AS1组LEF1-AS1 mRNA的相对表达量(0.059±0.010)低于NC组(1.000±0.018),差异有统计学意义(t=40.824,P<0.01);CCK-8实验结果显示,HUVECs在处理24 h后si-LEF1-AS1组细胞的吸光度(A)值(0.103±0.004)少于NC组(0.129±0.006),在处理48 h后si-LEF1-AS1组细胞的A值(0.235±0.016)也少于NC组(0.431±0.018),在处理72 h后si-LEF1-AS1组细胞的A值(0.454±0.029)也少于NC组(0.846±0.034);Transwell迁移实验结果显示si-LEF1-AS1组迁移到小室下侧的相对细胞数(0.446±0.025)少于NC组(1.000±0.046),差异有统计学意义(t=17.120,P<0.01);血管形成实验结果显示si-LEF1-AS1组的总成管长度(3152.333±200.959)少于NC组(13263.667±247.823),差异有统计学意义(t=54.891,P<0.01)。结论沉默长链非编码RNA LEF1-AS1抑制HUVEC细胞的增殖、迁移及血管生成能力。

Objective To investigate the effects of long non-coding RNA lymphoenhancer binding factor 1 antisense RNA1(LEF1-AS1)gene silencing on the proliferation,migration and angiogenesis of human umbilical vein endothelial cells(HUVECs).Methods HUVECs were obtained from american type culture collection(ATCC).The expression of LEF1-AS1 gene was silenced in huvecs by RNA interference.The cells were divided into control group and experimental group,namely NC-small interfering RNA(siRNA)group and LEF1-AS1-sirna group.The cell counting kit-8(CCK-8)and Transwell assays were used to detect the changes of cell proliferation and migration,and the effect of LEF1-AS1 on angiogenesis was detected in vitro.The experimental data were expressed as±s.The independent-sample t-test was used to compare the two groups.Results RT-qPCR results showed that the relative expression of LEF1-AS1 mRNA in si-LEF1-AS1 group(0.059±0.010)was lower than that in NC group(1.000±0.018),and the difference was statistically significant(t=40.824,P<0.01).HUVECs had lower absorbance(A)values(0.103±0.004)in si-LEF1-AS1 group than in NC group(0.129±0.006)at 24 h after treatment.The A values(0.235±0.016)in si-LEF1-AS1 group were also lower than in NC group(0.431±0.018)at 48 h after treatment.The A values in si-LEF1-AS1 group(0.454±0.029)were lower than those in NC group(0.846±0.034)after 72 h of treatment.Transwell migration experiment showed that the relative number of cells migrating to the inferior chamber in SI-LEF1-AS1 group(0.446±0.025)was significantly less than that in NC group(1.000±0.046,t=17.120,P<0.01).The results of angiogenic experiment showed that the length of the adult tube in SI-LEF1-AS1 group(3152.333±200.959)was significantly shorter than that in NC group(13263.667±247.823,t=54.891,P<0.01).Conclusion Silencing LEF1-AS1 inhibits the proliferation,migration and angiogenesis of HUVECs.