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环状 RNA 有_circ_0000527 通过 miR-646/BCL-2 轴参与视网膜母细胞瘤细胞的增殖、侵袭和迁移。

Circular RNA has_circ_0000527 participates in proliferation, invasion and migration of retinoblastoma cells via miR-646/BCL-2 axis.

机构信息

Neuroscience Center, Shandong Sunshine Union Hospital Co., Ltd., Weifang, China.

Department of Ophthalmology, Heze Municipal Hospital, Heze, China.

出版信息

Cell Biochem Funct. 2020 Dec;38(8):1036-1046. doi: 10.1002/cbf.3535. Epub 2020 Apr 7.

Abstract

Retinoblastoma (RB) is one of the most common primary intraocular malignancies in children. Emerging researches have shown that circular RNAs (circRNAs) play critical roles in a variety of cancers. As a novel circRNA, the function of circ_0000527 in RB remains unknown. In this work, expression level of circ_0000527 and miR-646 in RB tissues and cell lines were detected by quantitative real-time polymerase chain reaction (qRT-PCR). RB cell lines (SO-Rb50 and WERI-Rb-1) were used as cell models in functional experiments. CCK-8 assay, TUNEL assay and transwell assay were employed to detect the biological influence of circ_0000527 and miR-646 on cancer cells in vitro. qRT-PCR, luciferase reporter assay, RIP assay and western blot were used for exploring the interactions among circ_0000527, miR-646 and BCL-2. It was demonstrated that expression level of circ_0000527 in RB samples was significantly up-regulated compared to normal tissues, while miR-646 was markedly down-regulated. Overexpression of circ_0000527 promoted the viability, migration and invasion of RB cells, while miR-646 transfection had the opposite effects. Circ_0000527 sponged miR-646 to regulate the expression of BCL-2. In conclusion, circ_0000527 could promote the development of RB by indirectly modulating BCL-2 via absorbing miR-646. SIGNIFICANCE OF THE STUDY: Expression level of circ_0000527 in RB samples was significantly up-regulated compared to normal tissues, while miR-646 was markedly down-regulated. Overexpression of circ_0000527 promoted the viability, migration and invasion of RB cells, while miR-646 transfection had the opposite effects. Circ_0000527 sponged miR-646 to regulate the expression of BCL-2.

摘要

视网膜母细胞瘤 (RB) 是儿童中最常见的原发性眼内恶性肿瘤之一。新兴研究表明,环状 RNA (circRNA) 在多种癌症中发挥着关键作用。作为一种新型的 circRNA,circ_0000527 在 RB 中的功能尚不清楚。在这项工作中,通过实时定量聚合酶链反应 (qRT-PCR) 检测 RB 组织和细胞系中 circ_0000527 和 miR-646 的表达水平。使用 RB 细胞系 (SO-Rb50 和 WERI-Rb-1) 作为细胞模型进行功能实验。CCK-8 测定、TUNEL 测定和 Transwell 测定用于检测 circ_0000527 和 miR-646 对体外癌细胞的生物学影响。qRT-PCR、荧光素酶报告基因测定、RIP 测定和 Western blot 用于探索 circ_0000527、miR-646 和 BCL-2 之间的相互作用。结果表明,与正常组织相比,RB 样本中 circ_0000527 的表达水平显著上调,而 miR-646 的表达水平明显下调。circ_0000527 的过表达促进了 RB 细胞的活力、迁移和侵袭,而 miR-646 的转染则产生了相反的效果。circ_0000527 通过吸附 miR-646 来调节 BCL-2 的表达。总之,circ_0000527 通过间接调节 BCL-2 来促进 RB 的发展。

这项研究的意义在于

与正常组织相比,RB 样本中 circ_0000527 的表达水平显著上调,而 miR-646 的表达水平明显下调。circ_0000527 的过表达促进了 RB 细胞的活力、迁移和侵袭,而 miR-646 的转染则产生了相反的效果。circ_0000527 通过吸附 miR-646 来调节 BCL-2 的表达。

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