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SMAD6由DNM3OS-miR-134-5p轴正向调控，对视网膜母细胞瘤的细胞增殖、迁移和上皮-间质转化过程具有促进作用。

SMAD6, positively regulated by the DNM3OS-miR-134-5p axis, confers promoting effects to cell proliferation, migration and EMT process in retinoblastoma.

作者信息

Wang Hui, Ji Xiang

机构信息

Ophthalmology, The People's Hospital of Jiaozuo City, 267 Jiefang Middle Road, Jiaozuo City, 454150 Henan Province China.

出版信息

Cancer Cell Int. 2020 Jan 22;20:23. doi: 10.1186/s12935-020-1103-8. eCollection 2020.

DOI:10.1186/s12935-020-1103-8
PMID:31992960
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6977187/
Abstract

BACKGROUND

Retinoblastoma (RB) is acknowledged to be the commonest intraocular malignancy in infants and children and the outcome of RB patients is unfavorable due to limited early diagnosis and effective therapy. SMAD family member 6 (SMAD6) has been reported in the initiation and progression of human cancers by acting as a biological participant. However, the role of SMAD6 in RB has not been illustrated yet.

METHODS

The expression of SMAD6 mRNA, miR-134-5p and DNM3OS was measured by RT-qPCR. SMAD6 protein levels were measured by western blot. The effects of SMAD6 depletion on RB cells were analyzed using CCK-8 and transwell assays. The key proteins related to epithelial-mesenchymal transition (EMT) was determined by western blot. The localization of DNM3OS was detected by nuclear/cytoplasmic assay. In addition, the direct interaction between miR-134-5p and SMAD6 or DNM3OS was confirmed with the application of dual-luciferase reporter assay.

RESULTS

SMAD6 was upregulated in RB tissue samples and cell lines, and silencing SMAD6 suppressed cell proliferation, migration and EMT in RB. Mechanically, SMAD6 was positively regulated by lncRNA DNM3OS through competitively interacting with miR-134-5p. DNM3OS contributed to RB progression by SMAD6-mediated manner.

CONCLUSIONS

This research unmasked a novel DNM3OS/miR-134-5p/SMAD6 pathway in RB, which might make contribution to treatment of RB.

摘要

背景

视网膜母细胞瘤（RB）被认为是婴幼儿最常见的眼内恶性肿瘤，由于早期诊断和有效治疗有限，RB患者的预后不佳。据报道，SMAD家族成员6（SMAD6）作为生物学参与者在人类癌症的发生和发展中发挥作用。然而，SMAD6在RB中的作用尚未阐明。

方法

通过RT-qPCR检测SMAD6 mRNA、miR-134-5p和DNM3OS的表达。通过蛋白质免疫印迹法检测SMAD6蛋白水平。使用CCK-8和Transwell实验分析SMAD6缺失对RB细胞的影响。通过蛋白质免疫印迹法测定与上皮-间质转化（EMT）相关的关键蛋白。通过核/质实验检测DNM3OS的定位。此外，应用双荧光素酶报告基因实验证实miR-134-5p与SMAD6或DNM3OS之间的直接相互作用。

结果

SMAD6在RB组织样本和细胞系中上调，沉默SMAD6可抑制RB细胞的增殖迁移和EMT。机制上，lncRNA DNM3OS通过与miR-134-5p竞争性相互作用正向调节SMAD6。DNM3OS以SMAD6介导的方式促进RB进展。

结论

本研究揭示了RB中一条新的DNM3OS/miR-134-5p/SMAD6通路，这可能为RB的治疗做出贡献。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cba/6977187/ed9acecf92cb/12935_2020_1103_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cba/6977187/5588d29b5d26/12935_2020_1103_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cba/6977187/70e394f13df0/12935_2020_1103_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cba/6977187/9df7123df9ac/12935_2020_1103_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cba/6977187/e12950252b41/12935_2020_1103_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cba/6977187/ed9acecf92cb/12935_2020_1103_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cba/6977187/5588d29b5d26/12935_2020_1103_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cba/6977187/70e394f13df0/12935_2020_1103_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cba/6977187/9df7123df9ac/12935_2020_1103_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cba/6977187/e12950252b41/12935_2020_1103_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cba/6977187/ed9acecf92cb/12935_2020_1103_Fig5_HTML.jpg

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