Department of Ophthalmology, The Fourth People's Hospital of Shenyang, Shenyang City, Liaoning Province, China.
Department of Ophthalmology, The Fourth People's Hospital of Shenyang, Shenyang City, Liaoning Province, China.
Exp Eye Res. 2021 May;206:108529. doi: 10.1016/j.exer.2021.108529. Epub 2021 Mar 4.
Retinoblastoma (RB) is an intraocular malignancy that occurs in children. Circular RNAs (circRNAs) have been confirmed to play an essential role in tumorigenesis and development. This study aimed to ascertain the role and potential mechanism of hsa_circ_0099198 in RB. The levels of circ_0099198, microRNA-1287 (miR-1287) and low-density lipoprotein receptor-related protein 6 (LRP6) were determined by real-time quantitative polymerase chain reaction and Western blot. Cell proliferation was assessed by colony formation assay. Cell cycle arrest and apoptosis were evaluated by flow cytometry. Cell migration and invasion were tested using transwell assay. The activity of caspase-3/caspase-9 was examined with commercial kits. The interaction among circ_0099198, miR-1287 and LRP6 were verified by dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay or RNA pull-down assay. Xenograft experiment was used to assess tumor growth in vivo. circ_0099198 and LRP6 levels were increased, while miR-1287 level was reduced in RB cells. circ_0099198 silencing suppressed proliferation and metastasis and expedited cell cycle arrest and apoptosis in Y79 and So-RB50 cells. In addition, depletion of circ_0099198 inhibited RB cell progression via regulating miR-1287/LRP6 axis. Moreover, knockdown of circ_0099198 blocked the growth of xenograft tumors. circ_0099198 contributed to RB progression by sponging miR-1287 and up-regulating LRP6, which provided novel biomarkers for RB therapy.
视网膜母细胞瘤(RB)是一种发生在儿童中的眼内恶性肿瘤。环状 RNA(circRNA)已被证实在肿瘤发生和发展中发挥重要作用。本研究旨在确定 hsa_circ_0099198 在 RB 中的作用和潜在机制。通过实时定量聚合酶链反应和 Western blot 测定 circ_0099198、microRNA-1287(miR-1287)和低密度脂蛋白受体相关蛋白 6(LRP6)的水平。通过集落形成试验评估细胞增殖。通过流式细胞术评估细胞周期停滞和细胞凋亡。通过 Transwell 测定评估细胞迁移和侵袭。使用商业试剂盒检测 caspase-3/caspase-9 的活性。通过双荧光素酶报告基因测定、RNA 免疫沉淀(RIP)测定或 RNA 下拉测定验证 circ_0099198、miR-1287 和 LRP6 之间的相互作用。通过异种移植实验评估体内肿瘤生长。在 RB 细胞中,circ_0099198 和 LRP6 水平升高,而 miR-1287 水平降低。circ_0099198 沉默抑制 Y79 和 So-RB50 细胞的增殖和转移,并加速细胞周期停滞和凋亡。此外,circ_0099198 的耗竭通过调节 miR-1287/LRP6 轴抑制 RB 细胞进展。此外,circ_0099198 的敲低阻止了异种移植肿瘤的生长。circ_0099198 通过海绵吸附 miR-1287 和上调 LRP6 促进 RB 进展,为 RB 治疗提供了新的生物标志物。