干扰DBH-AS1上调miR-149抑制子宫内膜癌细胞增殖、迁移与侵袭  

Inhibition of the proliferation, migration and invasion of endometrial cancer cells through up-regulating miR-149 by interfering with DBH-AS1

作　　者：徐海霞[1] 伊碧霞[1] 胡娥 XU Haixia;YI Bixia;HU E(Jinhua People's Hospital,Department of Obstetrics and Gynecology,Jinhua,Zhejiang 321000,China)

机构地区：[1]金华市人民医院妇产科,浙江金华321000

出　　处：《中国优生与遗传杂志》2021年第11期1538-1543,共6页Chinese Journal of Birth Health & Heredity

基　　金：2021年度公益类金华市科学技术研究计划项目(2021-4-220)。

摘　　要：目的研究干扰长链非编码RNA(lncRNA)DBH-AS1是否通过上调微小RNA-149(miR-149)影响子宫内膜癌细胞的增殖、迁移与侵袭。方法将子宫内膜癌细胞RL95-2分为si-DBH-AS1组(转染DBH-AS1小干扰RNA si-DBH-AS1)、si-NC组(转染小干扰RNA阴性对照si-NC)、si-DBH-AS1+anti-miR-149组(同时转染si-DBH-AS1和miR-149抑制剂anti-miR-149)、si-DBH-AS1+anti-miR-NC组(同时转染si-DBH-AS1和miR-149抑制剂阴性对照anti-miR-NC)。采用荧光定量PCR(qRT-PCR)检测各组细胞中DBH-AS1和miR-149的表达,噻唑蓝(MTT)法检测细胞活性,流式细胞术检测细胞周期,Transwell检测迁移、侵袭细胞数目,免疫印迹实验(Western blot)检测细胞核相关抗原Ki67、N-钙黏蛋白(N-cadherin)、E-钙黏蛋白(E-cadherin)蛋白的表达。生物信息学预测与双荧光素酶实验DBH-AS1与miR-149的靶向结合。结果24例子宫内膜癌组织中DBH-AS1的表达水平显著升高,miR-149的表达水平显著降低,二者呈显著负相关(P<0.05)。与si-NC组相比,si-DBH-AS1组子宫内膜癌细胞的存活率、S期细胞比例、迁移细胞数、侵袭细胞数、Ki67、N-cadherin蛋白表达水平显著减少,miR-149表达水平、G0-G1期细胞比例、E-cadherin的蛋白表达水平显著增加(P<0.05)。与si-DBH-AS1+anti-miR-NC组相比,si-DBH-AS1+anti-miR-149组显著提高子宫内膜癌细胞的存活率、S期细胞比例、迁移细胞数、侵袭细胞数、Ki67、N-cadherin蛋白表达水平,显著降低miR-149表达水平、G0-G1期细胞比例、E-cadherin的蛋白表达水平(P<0.05)。DBH-AS1可以靶向调控miR-149的表达。结论干扰DBH-AS1可以上调miR-149的表达,抑制子宫内膜癌细胞的增殖、迁移与侵袭。Objective To investigate whether interfering long-chain non-coding RNA(lncRNA) DBH-AS1 affects the proliferation, migration and invasion of endometrial cancer cells by up-regulating microRNA-149(miR-149). Methods Endometrial cancer cell RL95-2 was divided into si-DBH-AS1 group(transfected with DBH-AS1 small interfering RNA si-DBH-AS1), si-NC group(transfected with small interfering RNA negative control si-NC), si-DBH-AS1+anti-miR-149 group(simultaneous transfection of si-DBH-AS1 and miR-149 inhibitor anti-miR-149), si-DBH-AS1+anti-miR-NC group(simultaneous transfection si-DBH-AS1 and miR-149 inhibitor negative control anti-miR-NC). Quantitative real-time PCR(qRT-PCR) was used to detect the expression of DBH-AS1 and miR-149 in each group of cells, thiazole blue(MTT) method was applied to assay cell activity, flow cytometry determined cell cycle, Transwell was performed to evaluate the number of migrating and invading cells, and Western blot experiments analyzed the expression of Ki67, N-cadherin, and E-cadherin protein. Bioinformatics prediction and dual luciferase experiment DBH-AS1 and miR-149 targeted binding. Results The expression level of DBH-AS1 in 24 endometrial cancer tissues significantly enhanced, the expression level of miR-149 significantly reduced, and the two were significantly negatively correlated(P<0.05). Compared with the si-NC group, the survival rate, the proportion of S-phase cells, the number of migrating cells and invasive cells, the expression of Ki67 and N-cadherin protein in the si-DBH-AS1 group remarkably reduced. miR-149 expression level, G0-G1 phase cell ratio, and E-cadherin protein expression level increased significantly(P<0.05). Compared with the si-DBH-AS1+anti-miR-NC group, the si-DBH-AS1+anti-miR-149 group greatly improved the survival rate of endometrial cancer cells, the proportion of S-phase cells, the number of migrated cells and the invasion cells, Ki67, N-cadherin protein expression level, while dramatically reduced miR-149 expression level, G0-G1 phase cell ratio, E-cadh

关 键 词：子宫内膜癌 DBH-AS1 miR-149 增殖 迁移 侵袭 

分 类 号：R737.33[医药卫生—肿瘤]