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长非编码 RNA LOXL1-AS1 通过海绵吸附 miR-589-5p 上调肾细胞癌中 CBX5 的表达。

Long non-conding RNA LOXL1-AS1 sponges miR-589-5p to up-regulate CBX5 expression in renal cell carcinoma.

机构信息

Department of Urology, The First Affiliated Hospital of Xinxiang Medical University, Weihui 453100, Henan, China.

Department of Urology, The Second Affiliated Hospital of Guangxi Medical University, Nanning 530000, Guangxi, China.

出版信息

Biosci Rep. 2020 Nov 27;40(11). doi: 10.1042/BSR20200212.

Abstract

BACKGROUND

Renal cell carcinoma (RCC) is a common malignant tumor that seriously endangers people's health. In recent years, long non-coding RNAs (lncRNAs) have been discovered to play vital roles in diverse cancers, including RCC. LncRNA lysyl oxidase like 1 antisense RNA 1 (LOXL1-AS1) has been found to exert carcinogenic functions in several cancers, but its role and mechanism in RCC have not been investigated.

METHODS

qRT-PCR was utilized for testing RNA expression and Western blot for protein expression in RCC tissues or cells. Then, we assessed cell function by conducting a series of functional experiments, such as 5-ethynyl-2'-deoxyuridine staining, colony formation, flow cytometry, JC-1, Western blot and transwell migration experiments. Following, RNA immunoprecipitation, pull down and luciferase reporter experiments were carried out to explore the regulatory mechanisms of LOXL1-AS1 in RCC.

RESULTS

LOXL1-AS1 was highly expressed in RCC tissues and cells. Moreover, knockdown of LOXL1-AS1 hampered RCC cell proliferation and migration. Importantly, miR-589-5p that was lowly expressed and worked as a tumor-inhibitor in RCC was found to bind with LOXL1-AS1. Furthermore, chromobox 5 (CBX5) targeted by miR-589-5p could expedite cell proliferation and migration in RCC. Finally, overexpressed CBX5 or inhibited miR-589-5p reversed the repressive impacts of silenced LOXL1-AS1 on RCC malignant phenotypes.

CONCLUSIONS

LncRNA LOXL1-AS1 sequestered miR-589-5p to augment CBX5 expression in RCC cells, opening a new way for potential development in RCC treatment.

摘要

背景

肾细胞癌(RCC)是一种严重危害人们健康的常见恶性肿瘤。近年来,长链非编码 RNA(lncRNA)被发现在包括 RCC 在内的多种癌症中发挥重要作用。赖氨酸氧化酶样 1 反义 RNA 1(LOXL1-AS1)已被发现在几种癌症中发挥致癌作用,但它在 RCC 中的作用和机制尚未得到研究。

方法

qRT-PCR 用于检测 RCC 组织或细胞中的 RNA 表达,Western blot 用于检测蛋白质表达。然后,我们通过一系列功能实验评估细胞功能,如 5-乙炔基-2'-脱氧尿苷染色、集落形成、流式细胞术、JC-1、Western blot 和 Transwell 迁移实验。随后,进行 RNA 免疫沉淀、下拉和荧光素酶报告基因实验,以探讨 LOXL1-AS1 在 RCC 中的调控机制。

结果

LOXL1-AS1 在 RCC 组织和细胞中高表达。此外,敲低 LOXL1-AS1 抑制了 RCC 细胞的增殖和迁移。重要的是,miR-589-5p 在 RCC 中低表达并发挥肿瘤抑制作用,被发现与 LOXL1-AS1 结合。此外,miR-589-5p 的靶基因 CBX5 可促进 RCC 细胞的增殖和迁移。最后,过表达 CBX5 或抑制 miR-589-5p 逆转了沉默 LOXL1-AS1 对 RCC 恶性表型的抑制作用。

结论

LncRNA LOXL1-AS1 通过与 miR-589-5p 结合来增强 RCC 细胞中 CBX5 的表达,为 RCC 的潜在治疗方法开辟了新的途径。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d1d/7670581/bcb844c013bc/bsr-40-bsr20200212-g1.jpg

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