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circ_0079586表达升高通过与miR-183-5p/信号通路相互作用预测胶质瘤预后不良并加速其进展。

Elevation in the Expression of circ_0079586 Predicts Poor Prognosis and Accelerates Progression in Glioma via Interactions with the miR-183-5p/ Signaling Pathway.

作者信息

Chen Jingyu, Liu Tianyi, Wang Hui, Wang Zhipeng, Lv Yanju, Zhao Yuying, Yang Ning, Yuan Xueli

机构信息

Department of Oncology, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang Province 150001, People's Republic of China.

Department of Pathology, The Second Hospital of Harbin Medical University, Harbin, Heilongjiang Province 150086, People's Republic of China.

出版信息

Onco Targets Ther. 2020 Jun 8;13:5135-5143. doi: 10.2147/OTT.S234758. eCollection 2020.

Abstract

PURPOSE

Glioma (GM) usually presents with an aggressive behavior and has a poor survival outcome. The abnormal expression of circular RNAs (circRNAs) has already been detected in GM, and circ_0079586 was found to have an increased expression in GM tumors.

PATIENTS AND METHODS

We assessed the differences in the expression of circ_0079586 in GM tissues (N=60) and cell lines (N=5) using qRT-PCR. The clinical value of circ_0079586 was measured by Fisher's exact test and Kaplan-Meier and Cox regression analyses. Circ_0079586 siRNA and vector were transfected into LN229 and U251 cells, respectively, and the transfection was verified by qRT-PCR. Cell growth was evaluated by cell counting kit-8 (CCK-8). Cell apoptosis was measured using flow cytometric assay. Cell metastatic properties were measured by wound scratch and transwell experiments. Subcellular fractionation was used to identify the location of circ_0079586. Dual-luciferase reporter test was utilized to confirm the interaction between miR-183-5p and circ_0079586/ 3'-UTR.

RESULTS

The expression of circ_0079586 was elevated in GM samples and cells and correlated with the clinical severity and unfavorable prognosis of the patients. The elevated expression of circ_0079586 led to an increase in cell growth, migration and invasion but inhibited apoptosis in U251 cells, whereas its down-regulation reversed these effects in the LN229 cells. Mechanistically, we found circ_0079586 to be primarily located in the cytoplasm of GM cells. Furthermore, circ_0079586 could act as a sponge for miR-183-5p and elevate expression at the posttranscriptional level.

CONCLUSION

In summary, circ_0079586 was found to be up-regulated in GM that increased the proliferation, migration and invasion in GM cells via interaction with the miR-183-5p/ axis. We anticipate that our study would provide newer insights into the mechanism and treatment of GM.

摘要

目的

胶质瘤(GM)通常具有侵袭性,生存预后较差。已在GM中检测到环状RNA(circRNA)的异常表达,且发现circ_0079586在GM肿瘤中表达增加。

患者与方法

我们使用qRT-PCR评估了circ_0079586在GM组织(N = 60)和细胞系(N = 5)中的表达差异。通过Fisher精确检验、Kaplan-Meier分析和Cox回归分析来测定circ_0079586的临床价值。将circ_0079586 siRNA和载体分别转染到LN229和U251细胞中,并通过qRT-PCR验证转染情况。使用细胞计数试剂盒-8(CCK-8)评估细胞生长。通过流式细胞术检测细胞凋亡。通过伤口划痕实验和Transwell实验测定细胞转移特性。采用亚细胞分级分离法确定circ_0079586的位置。利用双荧光素酶报告基因检测来确认miR-183-5p与circ_0079586/3'-UTR之间的相互作用。

结果

circ_0079586在GM样本和细胞中的表达升高,且与患者的临床严重程度和不良预后相关。circ_0079586的表达升高导致U251细胞的生长、迁移和侵袭增加,但抑制了细胞凋亡,而其下调则在LN229细胞中逆转了这些效应。从机制上讲,我们发现circ_0079586主要位于GM细胞的细胞质中。此外,circ_0079586可以作为miR-183-5p的海绵,并在转录后水平提高其表达。

结论

总之,发现circ_0079586在GM中上调,其通过与miR-183-5p相互作用增加了GM细胞的增殖、迁移和侵袭。我们预计我们的研究将为GM的机制和治疗提供新的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e9a7/7292368/285a1b0f6d6b/OTT-13-5135-g0001.jpg

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