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环状 RNA 通过 miR-361-3p/ADAM19 轴促进视网膜母细胞瘤进展。

Circular RNA has_circ_0000034 accelerates retinoblastoma advancement through the miR-361-3p/ADAM19 axis.

机构信息

Department of Ophthalmology, The Fourth People's Hospital of Shenyang, No. 20 Huanghe South Street, Huanggu District, Shenyang, 110031, Liaoning, China.

出版信息

Mol Cell Biochem. 2021 Jan;476(1):69-80. doi: 10.1007/s11010-020-03886-5. Epub 2020 Aug 25.

Abstract

Retinoblastoma (RB) is an intraocular malignancy that mainly occurs in infants and young children under 5 years of age. Circular RNA hsa_circ_0000034 (circ_0000034) was reported to be upregulated in RB tissues. Nevertheless, the function and mechanism of circ_0000034 in RB are unclear. Expression of circ_0000034, microRNA-361-3p (miR-361-3p), and a disintegrin and metalloproteinase 19 (ADAM19) was examined via quantitative real-time polymerase chain reaction (qRT-PCR). Cell viability, migration, invasion, and apoptosis were determined though Cell Counting Kit-8 (CCK-8), transwell, or flow cytometry assays. Caspase-3 activity was detected using a caspase-3 activity assay kit. Some protein levels were examined using Western blot analysis. Dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay, or RNA pull-down assay were performed to verify the relationship between circ_0000034 or ADAM19 and miR-361-3p. The function of circ_0000034 in vivo was confirmed via animal experiment. We verified that circ_0000034 expression was elevated in RB tissues and cells. Circ_0000034 silencing reduced RB growth in vivo, repressed viability, migration, invasion, and EMT, and induced apoptosis of RB cells in vitro. Circ_0000034 acted as a sponge for miR-361-3p, which targeted ADAM19 in RB cells. Furthermore, the inhibition of miR-361-3p restored circ_0000034 knockdown-mediated impacts on viability, migration, invasion, apoptosis, and EMT of RB cells. Moreover, ADAM19 overexpression abolished the influence of miR-361-3p mimic on viability, migration, invasion, apoptosis, and EMT of RB cells. Circ_0000034 expedited RB progression through upregulating ADAM19 via sponging miR-361-3p, which indicated that circ_0000034 might a target for RB therapy.

摘要

视网膜母细胞瘤 (RB) 是一种眼内恶性肿瘤,主要发生在 5 岁以下的婴儿和幼儿中。环状 RNA hsa_circ_0000034 (circ_0000034) 在 RB 组织中被报道上调。然而,circ_0000034 在 RB 中的功能和机制尚不清楚。通过实时定量聚合酶链反应 (qRT-PCR) 检测 circ_0000034、微小 RNA-361-3p (miR-361-3p) 和解整合素金属蛋白酶 19 (ADAM19) 的表达。通过细胞计数试剂盒-8 (CCK-8)、Transwell 或流式细胞术测定细胞活力、迁移和侵袭。使用 caspase-3 活性测定试剂盒检测 caspase-3 活性。通过 Western blot 分析检测某些蛋白水平。通过双荧光素酶报告基因检测、RNA 免疫沉淀 (RIP) 检测或 RNA 下拉检测验证 circ_0000034 或 ADAM19 与 miR-361-3p 的关系。通过动物实验证实 circ_0000034 在体内的功能。我们验证了 circ_0000034 在 RB 组织和细胞中的表达升高。circ_0000034 沉默减少了 RB 在体内的生长,抑制了 RB 细胞的活力、迁移、侵袭和 EMT,并诱导了体外 RB 细胞的凋亡。circ_0000034 作为 miR-361-3p 的海绵体,在 RB 细胞中靶向 ADAM19。此外,抑制 miR-361-3p 恢复了 circ_0000034 敲低对 RB 细胞活力、迁移、侵袭、凋亡和 EMT 的影响。此外,ADAM19 的过表达消除了 miR-361-3p 模拟对 RB 细胞活力、迁移、侵袭、凋亡和 EMT 的影响。Circ_0000034 通过海绵 miR-361-3p 上调 ADAM19 加速 RB 进展,表明 circ_0000034 可能成为 RB 治疗的靶点。

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