Department of Ophthalmology, Ningbo Eye Hospital, Ningbo, Zhejiang, China (mainland).
Med Sci Monit. 2020 Jun 15;26:e920886. doi: 10.12659/MSM.920886.
BACKGROUND Long noncoding RNA (lncRNA) acts as key regulator in human cancers, including retinoblastoma. However, the function of LINC00152 remains largely unknown in retinoblastoma. Thus, this study aimed to explore the role and molecular mechanisms of LINC00152 in retinoblastoma. MATERIAL AND METHODS The real-time quantitative polymerase chain reaction (RT-qPCR) was used to quantify the expression levels of LINC00152, miR-613 and yes-associated protein 1 (YAP1). The target genes of LINC00152 and miR-613 were identified by dual-luciferase reporter analysis, RNA immunoprecipitation (RIP) and RNA pulldown assays. The viability, apoptosis, and invasion of retinoblastoma cells were assessed by Cell Counting Kit-8, flow cytometry, and Transwell assays, respectively. In addition, western blot was used to test the protein expression in retinoblastoma cells or tissues. Cell sensitivity to carboplatin and adriamycin was analyzed by computing IC₅₀ value. The effects of LINC00152 silencing in vivo were measured by a xenograft experiment. RESULTS LINC00152 was obviously upregulated, while miR-613 was decreased in retinoblastoma tissues and cells. MiR-613, a target of LINC00152, was negatively regulated by LINC00152. Functional experiment further illustrated that silencing of LINC00152 evidently repressed proliferation, invasion, and autophagy while reinforced apoptosis of retinoblastoma cells, besides, retinoblastoma cells were more sensitive to carboplatin and adriamycin after knockdown of LINC00152. Importantly, knockdown of LINC00152-induced effects on retinoblastoma cells could be overturned by introducing miR-613 inhibitor. Downregulation of miR-613 abolished silencing of YAP1-effects on proliferation, apoptosis, invasion, autophagy, and chemoresistance of retinoblastoma cells. The results of the xenograft experiment indicated that LINC00152 silencing impeded tumor growth in vivo. CONCLUSIONS Mechanistically, LINC00152 enhanced the aggressiveness of retinoblastoma and boosted carboplatin and adriamycin resistance by regulating YAP1 by sponging miR-613 in human retinoblastoma.
长链非编码 RNA（lncRNA）在人类癌症中发挥关键调节作用，包括视网膜母细胞瘤。然而，LINC00152 在视网膜母细胞瘤中的作用仍知之甚少。因此，本研究旨在探讨 LINC00152 在视网膜母细胞瘤中的作用和分子机制。
实时定量聚合酶链反应（RT-qPCR）用于量化 LINC00152、miR-613 和 yes 相关蛋白 1（YAP1）的表达水平。通过双荧光素酶报告分析、RNA 免疫沉淀（RIP）和 RNA 下拉实验鉴定 LINC00152 和 miR-613 的靶基因。通过细胞计数试剂盒-8、流式细胞术和 Transwell 分析分别评估视网膜母细胞瘤细胞的活力、凋亡和侵袭。此外，Western blot 用于检测视网膜母细胞瘤细胞或组织中的蛋白质表达。通过计算 IC₅₀ 值分析细胞对卡铂和阿霉素的敏感性。通过异种移植实验测量 LINC00152 沉默的体内效果。
LINC00152 在视网膜母细胞瘤组织和细胞中明显上调，而 miR-613 下调。LINC00152 的靶基因 miR-613 受 LINC00152 的负调控。功能实验进一步表明，沉默 LINC00152 可显著抑制视网膜母细胞瘤细胞的增殖、侵袭和自噬，同时增强细胞凋亡，此外，敲低 LINC00152 后，视网膜母细胞瘤细胞对卡铂和阿霉素的敏感性增加。重要的是，引入 miR-613 抑制剂可逆转 LINC00152 敲低对视网膜母细胞瘤细胞的影响。下调 miR-613 可消除沉默 YAP1 对视网膜母细胞瘤细胞增殖、凋亡、侵袭、自噬和化疗耐药性的影响。异种移植实验的结果表明，LINC00152 沉默可抑制体内肿瘤生长。
从机制上讲，LINC00152 通过海绵 miR-613 调节 YAP1，增强了人视网膜母细胞瘤的侵袭性，并增强了对卡铂和阿霉素的耐药性。
