环状 RNA-E2F3 通过作为 miR-204-5p 的 ceRNA 调控 ROCK1 表达促进视网膜母细胞瘤的增殖、转移和抑制凋亡。
Circ-E2F3 acts as a ceRNA for miR-204-5p to promote proliferation, metastasis and apoptosis inhibition in retinoblastoma by regulating ROCK1 expression.
机构信息
Department of Ophthalmology, Guiping People's Hospital, Guiping 537200, Guangxi, China.
Department of Ophthalmology, Liuzhou People's Hospital, Liuzhou 545006, Guangxi, China.
出版信息
Exp Mol Pathol. 2021 Jun;120:104637. doi: 10.1016/j.yexmp.2021.104637. Epub 2021 Apr 18.
BACKGROUND
Circular RNA (circRNA) plays an important role in the malignant progression of many tumors, including retinoblastoma (RB). However, the role and regulatory mechanism of circ-E2F3 in RB have not been fully elucidated.
METHODS
Quantitative real-time PCR was used to measure circ-E2F3, miR-204-5p and Rho-associated protein kinase 1 (ROCK1) expression. Cell proliferation, apoptosis and metastasis were monitored by MTT, colony formation, flow cytometry, transwell and wound healing assays. Dual-luciferase reporter assay was employed to verify the relationship between miR-204-5p and circ-E2F3 or ROCK1. ROCK1 protein expression was detected by western blot assay. Mice xenograft models were built to assess the role of circ-E2F3 on RB tumor growth.
RESULTS
Circ-E2F3 was upregulated in RB tissues and cells. Silencing of circ-E2F3 inhibited the proliferation, migration, invasion, and induced the apoptosis of RB cells in vitro, as well as reduced RB tumor growth in vivo. MiR-204-5p could be sponged by circ-E2F3, and its inhibitor reversed the suppressive effect of circ-E2F3 silencing on RB progression. In addition, ROCK1 was confirmed to interact with miR-204-5p. MiR-204-5p regulated RB progression by targeting ROCK1. Also, circ-E2F3 positively regulated ROCK1 expression by sponging miR-204-5p.
CONCLUSION
Circ-E2F3 functioned as a tumor promoter in RB through the miR-204-5p/ROCK1 axis.
背景
环状 RNA（circRNA）在包括视网膜母细胞瘤（RB）在内的许多肿瘤的恶性进展中发挥着重要作用。然而，circ-E2F3 在 RB 中的作用和调控机制尚未完全阐明。
方法
采用实时定量 PCR 检测 circ-E2F3、miR-204-5p 和 Rho 相关蛋白激酶 1（ROCK1）的表达。通过 MTT、集落形成、流式细胞术、Transwell 和划痕愈合实验监测细胞增殖、凋亡和转移。双荧光素酶报告实验验证 miR-204-5p 与 circ-E2F3 或 ROCK1 的关系。Western blot 检测 ROCK1 蛋白表达。构建小鼠异种移植模型评估 circ-E2F3 对 RB 肿瘤生长的作用。
结果
circ-E2F3 在 RB 组织和细胞中上调。沉默 circ-E2F3 抑制 RB 细胞的增殖、迁移、侵袭，并在体外诱导凋亡，同时减少体内 RB 肿瘤的生长。circ-E2F3 可以作为 miR-204-5p 的海绵，其抑制剂逆转了 circ-E2F3 沉默对 RB 进展的抑制作用。此外，ROCK1 被证实与 miR-204-5p 相互作用。miR-204-5p 通过靶向 ROCK1 调节 RB 进展。此外，circ-E2F3 通过海绵吸附 miR-204-5p 正向调节 ROCK1 表达。
结论
circ-E2F3 通过 miR-204-5p/ROCK1 轴在 RB 中发挥肿瘤促进作用。
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