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微小RNA-106b通过抑制ZBTB4蛋白的表达促进视网膜母细胞瘤细胞的增殖、迁移和侵袭。

MicroRNA-106b promotes the proliferation, migration and invasion of retinoblastoma cells by inhibiting the expression of ZBTB4 protein.

作者信息

Bu Wenjuan, Wang Yanhui, Min Xiangrong

机构信息

Department of Ophthalmology, Jining No. 1 People's Hospital, Jining, Shandong 272011, P.R. China.

Department of Fundus Surgery, Hebei Province Eye Hospital, Xingtai, Hebei 054000, P.R. China.

出版信息

Exp Ther Med. 2018 Dec;16(6):4537-4545. doi: 10.3892/etm.2018.6811. Epub 2018 Oct 1.

Abstract

The present study investigated the function of microRNA (miR)-106b in the proliferation, migration and invasion of retinoblastoma (RB) cells, and aimed to elucidate the underlying mechanism. A total of 56 patients with RB were enrolled in the present study. The expression of miR-106b in RB tissues was measured by reverse transcription quantitative polymerase chain reaction. After transfection with miR-106b mimics or miR-106b inhibitor, a Cell-Counting kit-8 assay was used to determine the proliferation of WERI-Rb-1 cells and a Transwell assay was employed to measure the migration and invasion of the cells. Western blot analysis was performed to determine the expression of zinc finger and BTB domain containing 4 (ZBTB4) protein. By silencing or overexpression of ZBTB4 protein, the biological functions of ZBTB4 in WERI-Rb-1 cells were studied. A dual luciferase reporter assay was performed to test whether ZBTB4 was a target gene of miR-106b. The expression of miR-106b in RB tissues was elevated and closely associated with the severity of the disease. Overexpression of miR-106b increased but inhibition of miR-106b expression decreased the proliferation, migration and invasion abilities of WERI-Rb-1 cells. In addition, overexpression of miR-106b decreased but inhibition of miR-106b expression increased ZBTB4 protein expression in WERI-Rb-1 cells. Similarly, overexpression of ZBTB4 reduced but inhibition of ZBTB4 expression promoted the proliferation, migration and invasion of WERI-Rb-1 cells. Finally, miR-106b regulated the expression of ZBTB4 by binding to the 3'-untranslated region of the ZBTB4 gene. The present study demonstrated that increased expression of miR-106b in RB tissues is positively associated with the metastasis and differentiation of RB cells. As an oncogene, miR-106b promotes the proliferation, migration and invasion of WERI-Rb-1 cells by inhibiting the expression of ZBTB4 protein.

摘要

本研究调查了微小RNA(miR)-106b在视网膜母细胞瘤(RB)细胞增殖、迁移和侵袭中的作用,旨在阐明其潜在机制。本研究共纳入56例RB患者。采用逆转录定量聚合酶链反应检测RB组织中miR-106b的表达。用miR-106b模拟物或miR-106b抑制剂转染后,使用细胞计数试剂盒-8法测定WERI-Rb-1细胞的增殖情况,采用Transwell法检测细胞的迁移和侵袭能力。进行蛋白质免疫印迹分析以测定含锌指和BTB结构域4(ZBTB4)蛋白的表达。通过沉默或过表达ZBTB4蛋白,研究ZBTB4在WERI-Rb-1细胞中的生物学功能。进行双荧光素酶报告基因检测以验证ZBTB4是否为miR-106b的靶基因。RB组织中miR-106b的表达升高,且与疾病严重程度密切相关。miR-106b的过表达增加了WERI-Rb-1细胞的增殖、迁移和侵袭能力,但抑制miR-106b的表达则降低了这些能力。此外,miR-106b的过表达降低了WERI-Rb-1细胞中ZBTB4蛋白的表达,但抑制miR-106b的表达则增加了ZBTB4蛋白的表达。同样,ZBTB4的过表达降低了WERI-Rb-1细胞的增殖、迁移和侵袭能力,但抑制ZBTB4的表达则促进了这些能力。最后,miR-106b通过与ZBTB4基因的3'非翻译区结合来调节ZBTB4的表达。本研究表明,RB组织中miR-106b表达的增加与RB细胞的转移和分化呈正相关。作为一种癌基因,miR-106b通过抑制ZBTB4蛋白的表达促进WERI-Rb-1细胞的增殖、迁移和侵袭。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d0c/6257475/e676f2aa706c/etm-16-06-4537-g00.jpg

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