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
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荆门市卫生计生委科研项目(No.202101021)


Effect of silencing LncRNA DLGAP1-AS2 on the proliferation, migration and invasion of human retinoblastoma
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Scientific Research Project of Jingmen Health and Family Planning Commission(No.202101021)

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    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。

    Abstract:

    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.

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李晖,汪明红,廖风玲,等.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.国际眼科杂志, 2022,22(6):904-910.

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  • 收稿日期:2021-07-01
  • 最后修改日期:2022-04-25
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  • 在线发布日期: 2022-05-30
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