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长链非编码RNA OIP5-AS1通过吸附miR-27a-3p激活TSC1表达来抑制多发性骨髓瘤进展。

Long non-coding RNA OIP5-AS1 suppresses multiple myeloma progression by sponging miR-27a-3p to activate TSC1 expression.

作者信息

Wang Yong, Wang Haibao, Ruan Jianwei, Zheng Wenbiao, Yang Zeyu, Pan Weiwei

机构信息

Department of Orthopaedic, Taizhou Municipal Hospital, No. 381, Zhongshan East Road, Jiaojiang District, Taizhou, 318000 Zhejiang China.

出版信息

Cancer Cell Int. 2020 May 7;20:155. doi: 10.1186/s12935-020-01234-7. eCollection 2020.

Abstract

BACKGROUND

Multiple myeloma (MM) is a prevalent hematological malignancy. Long noncoding RNAs are correlated with the development of MM. In this project, the function of lncRNA opa interacting protein 5-antisense 1 (OIP5-AS1) in MM and the potential mechanistic pathway were explored.

METHODS

The expression of OIP5-AS1, microRNA (miR)-27a-3p and tuberous sclerosis 1 (TSC1) was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) assay. Cell proliferation was assessed by Cell Counting Kit-8 (CCK-8) assay, colony formation assay and Bromodeoxyuridine (BrdU) staining. And cell apoptosis was evaluated by flow cytometry assay. Cell metastasis was assessed utilizing transwell assay. Western blot analysis was employed to detect protein level. The target relation between miR-27a-3p and OIP5-AS1 or TSC1 was confirmed via dual-luciferase reporter assay and RNA immunoprecipitation assay. Tumor xenograft assay was conducted to measure the function of OIP5-AS1 in vivo.

RESULTS

The expression levels of OIP5-AS1 and TSC1 were decreased in MM, whereas miR-27a-3p was upregulated. High level of OIP5-AS1 could predict favourable prognosis of MM patients. Overexpression of OIP5-AS1 inhibited cell viability, colony formation ability, migration and invasion, induced cell cycle arrest in G1 phase and apoptosis of MM cells in vitro as well as repressed tumorigenesis in vivo. MiR-27a-3p was a target of OIP5-AS1, and reversed the impact of OIP5-AS1 on MM cells. MiR-27a-3p directly targeted TSC1. Silencing of miR-27a-3p repressed MM progression by elevating TSC1 expression. OIP5-AS1 upregulated TSC1 by sponging miR-27a-3p.

CONCLUSION

OIP5-AS1 repressed multiple myeloma progression by regulating miR-27a-3p/TSC1 axis.

摘要

背景

多发性骨髓瘤(MM)是一种常见的血液系统恶性肿瘤。长链非编码RNA与MM的发生发展相关。本研究旨在探讨长链非编码RNA透明质酸结合蛋白5反义链1(OIP5-AS1)在MM中的作用及其潜在机制。

方法

采用定量实时聚合酶链反应(qRT-PCR)检测OIP5-AS1、微小RNA(miR)-27a-3p和结节性硬化症蛋白1(TSC1)的表达水平。采用细胞计数试剂盒-8(CCK-8)法、集落形成试验和溴脱氧尿苷(BrdU)染色评估细胞增殖。采用流式细胞术检测细胞凋亡。采用Transwell试验评估细胞转移。采用蛋白质印迹法检测蛋白水平。通过双荧光素酶报告基因试验和RNA免疫沉淀试验验证miR-27a-3p与OIP5-AS1或TSC1之间的靶向关系。通过肿瘤异种移植试验检测OIP5-AS1在体内的作用。

结果

MM患者中OIP5-AS1和TSC1的表达水平降低,而miR-27a-3p上调。高水平的OIP5-AS1可预测MM患者的良好预后。过表达OIP5-AS1可抑制体外MM细胞的活力、集落形成能力、迁移和侵袭,诱导细胞周期阻滞于G1期并促进细胞凋亡,同时抑制体内肿瘤发生。miR-27a-3p是OIP5-AS1的靶点,可逆转OIP5-AS1对MM细胞的影响。miR-27a-3p直接靶向TSC1。沉默miR-27a-3p可通过提高TSC1表达抑制MM进展。OIP5-AS1通过海绵吸附miR-27a-3p上调TSC1。

结论

OIP5-AS1通过调节miR-27a-3p/TSC1轴抑制多发性骨髓瘤进展。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a0c7/7206794/25066afa3c57/12935_2020_1234_Fig1_HTML.jpg

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