沉默LncRNA DLGAP1-AS2对人视网膜母细胞瘤增殖和迁移及侵袭的影响  

Effect of silencing LncRNA DLGAP1-AS2 on the proliferation,migration and invasion of human retinoblastoma

作　　者：李晖[1] 汪明红[1] 廖风玲[1] 刘国立[1] Hui Li;Ming-Hong Wang;Feng-Ling Liao;Guo-Li Liu(Department of Ophthalmology, the First People's Hospital of Jingmen City, Jingmen 448000, Hubei Province, China)

机构地区：[1]中国湖北省荆门市第一人民医院眼科,448000

出　　处：《国际眼科杂志》2022年第6期904-910,共7页International Eye Science

基　　金：荆门市卫生计生委科研项目(No.202101021)。

摘　　要：目的:探讨沉默LncRNA DLGAP1-AS2对人视网膜母细胞瘤HXO-Rb44增殖、迁移和侵袭的影响及其可能作用机制。方法:收集病理确诊且临床资料完整的视网膜母细胞瘤组织标本25例,同时选取因外伤摘除眼球的正常视网膜组织9例为对照;qRT-PCR法检测正常视网膜组织、视网膜母细胞瘤组织、人正常视网膜血管内皮细胞ACBRI-181、视网膜母细胞瘤细胞HXO-Rb44中DLGAP1-AS2、miR-1193的表达量;将si-NC、si-DLGAP1-AS2、miR-NC、miR-1193 mimic、si-DLGAP1-AS2与miR-1193 inhibitor(共转染)分别转染至HXO-Rb44细胞;双荧光素酶报告实验检测DLGAP1-AS2与miR-1193的靶向关系;CCK-8法、平板克隆形成实验与Transwell实验分别检测细胞增殖、集落形成数、迁移及侵袭;Western blot法检测E-cadherin、N-cadherin蛋白表达量。结果:视网膜母细胞瘤组织中DLGAP1-AS2的表达量高于正常视网膜组织(P<0.05),而miR-1193的表达量低于正常视网膜组织(P<0.05);HXO-Rb44细胞中DLGAP1-AS2的表达量高于ACBRI-181细胞(P<0.05),miR-1193的表达量低于ACBRI-181细胞(P<0.05);DLGAP1-AS2可靶向调控miR-1193的表达;转染si-DLGAP1-AS2或转染miR-1193 mimic可抑制HXO-Rb44细胞增殖、迁移及侵袭;共转染si-DLGAP1-AS2与miR-1193 inhibitor可降低转染si-DLGAP1-AS2对HXO-Rb44细胞增殖、迁移及侵袭的作用。结论:沉默DLGAP1-AS2可通过靶向调控miR-1193表达而抑制视网膜母细胞瘤细胞增殖、迁移及侵袭。AIM:To explore the effect of silencing LncRNA DLGAP1-AS2 on the proliferation,migration and invasion of human retinoblastoma HXO-Rb44 and its possible mechanism.METHODS:Twenty-five cases of retinoblastoma tissue specimens with complete clinical data and pathologically diagnosed were collected.At the same time,9 cases of normal retinal tissue from which the eyeball was removed due to trauma were selected as controls.The qRT-PCR method was used to detect the expression of DLGAP1-AS2 and miR-1193 in normal retinal tissue,retinoblastoma tissue,human normal retinal vascular endothelial cell ACBRI-181,and retinoblastoma cell HXO-Rb44.The si-NC,si-DLGAP1-AS2,miR-NC,miR-1193 mimic,si-DLGAP1-AS2 and miR-1193 inhibitor(co-transfected)were transfected into HXO-Rb44 cells.The dual luciferase reporter experiment was used to detect the targeting relationship between DLGAP1-AS2 and miR-1193.The CCK-8 method,plate clone formation experiment and Transwell experiment were used to detect cell proliferation,colony formation,migration and invasion.Western blot method was used to detect the expression of E-cadherin and N-cadherin protein.RESULTS:The expression of DLGAP1-AS2 in retinoblastoma tissue was higher than that of normal retinal tissue(P<0.05),while the expression of miR-1193 was lower than that of normal retinal tissue(P<0.05).The expression of DLGAP1-AS2 in HXO-Rb44 cells was higher than that of ACBRI-181 cells(P<0.05),and the expression of miR-1193 was lower than that of ACBRI-181 cells(P<0.05).DLGAP1-AS2 could target the expression of miR-1193.Transfection of si-DLGAP1-AS2 or miR-1193 mimic could inhibit the proliferation,migration and invasion of HXO-Rb44 cells.Co-transfection of si-DLGAP1-AS2 and miR-1193 inhibitor could reduce the effect of transfection of si-DLGAP1-AS2 on the proliferation,migration and invasion of HXO-Rb44 cells.CONCLUSION:Silencing DLGAP1-AS2 could inhibit the proliferation,migration and invasion of retinoblastoma cells through targeted regulation of miR-1193 expression.

关 键 词：人视网膜母细胞瘤 LncRNA DLGAP1-AS2 miR-1193 细胞增殖 迁移 侵袭 

分 类 号：R73[医药卫生—肿瘤]