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环状RNA_OTUD7A通过充当miR-431-5p的海绵上调FOXP1表达,以促进弥漫性大B细胞淋巴瘤的进展。

CircRNA_OTUD7A upregulates FOXP1 expression to facilitate the progression of diffuse large B-cell lymphoma via acting as a sponge of miR-431-5p.

作者信息

Liu Wei, Lei Lei, Liu Xiaoying, Ye Suiyan

机构信息

Department of Hematology, The First Hospital of Yulin, Yulin, Shaanxi, China.

Clinical Laboratory, The First Hospital of Yulin, No. 59 Yuxi Avenue, Yulin, 719000, Shaanxi, China.

出版信息

Genes Genomics. 2021 Jun;43(6):653-667. doi: 10.1007/s13258-021-01094-z. Epub 2021 Apr 8.

Abstract

BACKGROUND

A growing number of studies have shown that circular RNA (circRNA) is an important regulator molecule in cancer progression, but it has been poorly studied in diffuse large b-cell lymphoma (DLBCL).

OBJECTIVE

This study aimed to explore the role of circ_OTUD7A in DLBCL.

METHODS

Relative expression levels of circ_OTUD7A, microRNA (miR)-431-5p and forkhead box P1 (FOXP1) were determined by quantitative real-time PCR (qRT-PCR). The proliferation of cells was elevated by colony formation assay and MTT assay. Western blot (WB) analysis was employed to measure the protein levels of proliferation marker, epithelial-mesenchymal transition (EMT) markers, cyclin marker, apoptosis markers and FOXP1. Moreover, the apoptosis, cell cycle process, migration and invasion of cells were detected using flow cytometry and transwell assay, respectively. In addition, the interaction between miR-431-5p and circ_OTUD7A or FOXP1 was confirmed by dual-luciferase reporter assay.

RESULTS

Circ_OTUD7A was highly expressed in DLBCL, and its knockdown could inhibit DLBCL cell proliferation and metastasis, while promote cell cycle arrest and apoptosis. Similarly, FOXP1 also was upregulated in DLBCL, and its silencing could restrain the progression of DLBCL cells. Further experiments revealed that circ_OTUD7A could sponge miR-431-5p and miR-431-5p could target FOXP1. MiR-431-5p inhibitor could reverse the suppressive effect of circ_OTUD7A silencing on DLBCL progression, and FOXP1 overexpression also could reverse the inhibitory effect of miR-431-5p mimic on DLBCL progression.

CONCLUSION

Circ_OTUD7A promoted the progression of DLBCL by regulating the miR-431-5p/FOXP1 axis, which suggested that circ_OTUD7A might function as an oncogene in DLBCL.

摘要

背景

越来越多的研究表明,环状RNA(circRNA)是癌症进展中的重要调节分子,但在弥漫性大B细胞淋巴瘤(DLBCL)中的研究较少。

目的

本研究旨在探讨circ_OTUD7A在DLBCL中的作用。

方法

通过定量实时荧光定量PCR(qRT-PCR)检测circ_OTUD7A、微小RNA(miR)-431-5p和叉头框P1(FOXP1)的相对表达水平。采用集落形成试验和MTT试验检测细胞增殖情况。运用蛋白质免疫印迹(WB)分析检测增殖标志物、上皮-间质转化(EMT)标志物、细胞周期蛋白标志物、凋亡标志物及FOXP1的蛋白水平。此外,分别采用流式细胞术和Transwell试验检测细胞凋亡、细胞周期进程、迁移和侵袭情况。另外,通过双荧光素酶报告基因试验证实miR-431-5p与circ_OTUD7A或FOXP1之间的相互作用。

结果

Circ_OTUD7A在DLBCL中高表达,敲低其表达可抑制DLBCL细胞增殖和转移,同时促进细胞周期阻滞和凋亡。同样,FOXP1在DLBCL中也上调,沉默其表达可抑制DLBCL细胞进展。进一步实验表明,circ_OTUD7A可吸附miR-431-5p,且miR-431-5p可靶向FOXP1。miR-431-5p抑制剂可逆转circ_OTUD7A沉默对DLBCL进展的抑制作用,FOXP1过表达也可逆转miR-431-5p模拟物对DLBCL进展的抑制作用。

结论

Circ_OTUD7A通过调节miR-431-5p/FOXP1轴促进DLBCL进展,提示circ_OTUD7A可能在DLBCL中作为癌基因发挥作用。

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