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.
Cell Prolif. 2019 Sep;52(5):e12635. doi: 10.1111/cpr.12635. Epub 2019 Jul 23.
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.
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.
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.
Our study sheds light on the novel underlying mechanism of miR-191 in HCC, which may accelerate the development of cancer therapy.
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。
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。
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。
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。