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MicroRNA-191 通过 has_circ_0000204/miR-191/KLF6 轴促进肝癌细胞增殖。

MicroRNA-191 promotes hepatocellular carcinoma cell proliferation by has_circ_0000204/miR-191/KLF6 axis.

机构信息

Affiliated Hospital of Nanjing University of Chinese Medicine, Jiangsu Province Hospital of Chinese Medicine, Nanjing, China.

School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai, China.

出版信息

Cell Prolif. 2019 Sep;52(5):e12635. doi: 10.1111/cpr.12635. Epub 2019 Jul 23.

Abstract

OBJECTIVES

MicroRNAs are powerful regulators in hepatocellular carcinoma (HCC) tumorigenesis. MicoRNA-191 (miR-191) has been reported to play an important role in HCC, However, the regulatory mechanism is still unclear. In this study, we investigated the role of miR-191 in HCC and studied its underlying mechanisms of action.

MATERIALS AND METHODS

The expression of miR-191 in HCC tissues was determined by quantitative real-time PCR (qRT-PCR). The role of miR-191 in HCC cells was examined by using both in vitro and in vivo assays. Downstream targets of miR-191 were determined by qRT-PCR and Western blot analysis. Dual-luciferase assays were performed to validate the interaction between miR-191 and its targets.

RESULTS

The expression of miR-191 was significantly higher in HCC patients and a higher miR-191 expression predicted poorer prognosis. Analysis of The Cancer Genome Atlas data sets suggested that miR-191 positively correlated with cell cycle progression. Gain and loss of function assays showed that miR-191 promoted cell cycle progression and proliferation. Luciferase reporter assay showed that miR-191 directly targeted the 3'-untranslated region of KLF6 mRNA. Furthermore, circular RNA has_circ_0000204 could sponge with miR-191, resulting in inactivation of miR-191.

CONCLUSIONS

Our study sheds light on the novel underlying mechanism of miR-191 in HCC, which may accelerate the development of cancer therapy.

摘要

目的

微小 RNA(miRNA)是肝细胞癌(HCC)发生过程中的强大调控因子。有研究报道 miRNA-191(miR-191)在 HCC 中发挥重要作用,但调控机制尚不清楚。本研究旨在探讨 miR-191 在 HCC 中的作用及其作用机制。

材料和方法

采用实时定量 PCR(qRT-PCR)检测 HCC 组织中 miR-191 的表达。通过体外和体内实验研究 miR-191 在 HCC 细胞中的作用。采用 qRT-PCR 和 Western blot 分析检测 miR-191 的下游靶基因。采用双荧光素酶报告基因实验验证 miR-191 与其靶基因的相互作用。

结果

miR-191 在 HCC 患者中的表达明显升高,且 miR-191 高表达预示预后不良。分析癌症基因组图谱(TCGA)数据集提示 miR-191 与细胞周期进程呈正相关。Gain-和 loss-of-function 实验表明,miR-191 促进细胞周期进程和增殖。荧光素酶报告基因实验表明,miR-191 可直接靶向 KLF6 mRNA 的 3'-非翻译区。进一步研究发现,circRNA has_circ_0000204 可与 miR-191 结合,从而使 miR-191 失活。

结论

本研究揭示了 miR-191 在 HCC 中的新作用机制,可能有助于开发癌症治疗方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee1a/6797514/db667f1b8b90/CPR-52-e12635-g001.jpg

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