Yonglun Huang1, Boguang Xue1, Jianhui Pan1, Nian Shen2. 1. Department of Ophthalmology, Guiping People's Hospital, Guiping 537200, Guangxi, China. 2. Department of Ophthalmology, Liuzhou People's Hospital, Liuzhou 545006, Guangxi, China. Electronic address: Shennianzz@163.com.
Abstract
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.
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.