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。

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.