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Circ_0061140通过吸附微小RNA-1236促进膀胱癌转移。

Circ_0061140 promotes metastasis of bladder cancer through adsorbing microRNA-1236.

作者信息

Feng F, Chen A-P, Wang X-L, Wu G-L

机构信息

Department of Urology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, China.

出版信息

Eur Rev Med Pharmacol Sci. 2020 May;24(10):5310-5319. doi: 10.26355/eurrev_202005_21313.

Abstract

OBJECTIVE

The purpose of this study was to investigate the expression characteristics of circular RNA circ_0061140 in bladder cancer (BCa), and to further explore its effects on invasiveness and migration capacity of BCa cells, as well as its possible potential mechanism.

PATIENTS AND METHODS

Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to examine the expression level of circ_0061140 in tumor tissue samples and paracancerous ones collected from 42 patients with BCa, and the interplay between circ_0061140 level and the clinical indicators, as well as the prognosis of BCa patients were analyzed. Meanwhile, qRT-PCR was also used to verify circ_0061140 expression in BCa cell lines. In addition, a circ_0061140 knockdown model was constructed using Lentiviral in BCa cell lines, including T24 and 253j, and the effect of circ_0061140 on BCa cell functions and its underlying mechanisms were explored using Cell Counting Kit-8 (CCK-8), transwell, and cell wound healing assays.

RESULTS

qPCR results showed that the expression level of circ_0061140 in tumor tissues of BCa patients was remarkably higher than that in adjacent tissues, and the difference was statistically significant. Compared with patients with low expression of circ_0061140, patients with high expression of circ_0061140 had worse prognosis and higher incidence of lymph node or distant metastasis. Compared with those in the negative control group, the proliferation and invasion, as well as the metastasis ability of BCa cells in the sh-circ_0061140 group, were remarkably attenuated. In addition, bioinformatics and Luciferase reporter gene assay demonstrated that circ_0061140 can specifically bind to microRNA-1236. At the same time, the results of qPCR revealed that the expression levels of circ_0061140 and microRNA-1236 were negatively correlated in the tumor tissues of BCa patients. Finally, cell recovery experiment indicated that silencing microRNA-1236 reversed the impact of the knockdown of circ_0061140 on the ability of BCa cells to proliferate and invade, suggesting that the two may regulate each other.

CONCLUSIONS

Circ_0061140 level was found remarkably elevated in BCa tissues, as well as in cell lines, which was closely relevant to the incidence of lymph node or distant metastasis of BCa patients. In addition, circ_0061140 may enhance the proliferation rate and invasion ability of BCa cells through the modulation of microRNA-1236.

摘要

目的

本研究旨在探讨环状RNA circ_0061140在膀胱癌(BCa)中的表达特征,并进一步探究其对BCa细胞侵袭和迁移能力的影响及其可能的潜在机制。

患者与方法

采用定量实时聚合酶链反应(qRT-PCR)检测42例BCa患者肿瘤组织样本及癌旁组织中circ_0061140的表达水平,分析circ_0061140水平与临床指标之间的相互关系以及BCa患者的预后。同时,运用qRT-PCR验证circ_0061140在BCa细胞系中的表达。此外,利用慢病毒在BCa细胞系(包括T24和253j)中构建circ_0061140敲低模型,并通过细胞计数试剂盒-8(CCK-8)、Transwell和细胞划痕愈合实验探究circ_0061140对BCa细胞功能及其潜在机制的影响。

结果

qPCR结果显示,BCa患者肿瘤组织中circ_0061140的表达水平显著高于相邻组织,差异具有统计学意义。与circ_0061140低表达患者相比,circ_0061140高表达患者预后较差,淋巴结或远处转移发生率更高。与阴性对照组相比,sh-circ_0061140组BCa细胞的增殖、侵袭及转移能力均显著减弱。此外,生物信息学和荧光素酶报告基因检测表明circ_0061140可特异性结合微小RNA-1236。同时,qPCR结果显示,BCa患者肿瘤组织中circ_0061140与微小RNA-1236的表达水平呈负相关。最后,细胞回复实验表明,沉默微小RNA-1236可逆转circ_0061140敲低对BCa细胞增殖和侵袭能力的影响,提示二者可能相互调节。

结论

发现circ_0061140水平在BCa组织及细胞系中显著升高,这与BCa患者淋巴结或远处转移的发生率密切相关。此外,circ_0061140可能通过调控微小RNA-1236增强BCa细胞的增殖率和侵袭能力。

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