Department of Infectious Diseases, The First Affiliated Hospital of Guangxi Medical University, Nanning, China.
Eur Rev Med Pharmacol Sci. 2020 Jan;24(2):647-654. doi: 10.26355/eurrev_202001_20041.
MicroRNAs (miRNAs) play a role in the pathogenesis of hepatocellular carcinoma (HCC). This study was designed to elucidate the role of microRNA-31 (miR-31) in HCC.
HuH7 cell lines were transfected with miR-31 mimic or miR-31 inhibitor to investigate the role of miR-31 in regulating interferon regulatory factor-1 (IRF-1). The mRNA and protein expression levels of IRF-1 were quantitatively detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot, respectively. Subsequently, Dual-Luciferase reporter assay was also performed.
The expression level of miR-31 was significantly up-regulated in HuH7 cells when compared with that in primary human hepatocytes (hHC). Dual-Luciferase reporter assay indicated that IRF-1 was the direct target of miR-31. The expression levels of IRF-1 were decreased in HuH7 and HepG2 cell lines. IRF-1 was negatively correlated with miR-31 in HCC tissues and paired adjacent tissues. The expression level of miR-31 was inversely correlated with IRF-1. MiR-31 inhibitor up-regulated the expression levels of IRF-1 in HuH7 cells, whereas miR-31 mimic down-regulated the expression levels of IRF-1. Furthermore, the miR-31 mimic repressed IRF-1-3'UTR reporter activity, whereas the miR-31 inhibitor enhanced IRF-1-3'UTR reporter activity depending on the concentration of miR-31 mimic and miR-31 inhibitor.
These results indicated that miR-31 can regulate the expression level of IRF-1 in HCC, which probably provided novel theoretical evidence for the application of target miR-31 treatment of HCC.
微小 RNA（miRNA）在肝细胞癌（HCC）的发病机制中发挥作用。本研究旨在阐明 microRNA-31（miR-31）在 HCC 中的作用。
用 miR-31 模拟物或 miR-31 抑制剂转染 HuH7 细胞系，以研究 miR-31 在调节干扰素调节因子-1（IRF-1）中的作用。通过定量实时聚合酶链反应（qRT-PCR）和 Western blot 分别定量检测 IRF-1 的 mRNA 和蛋白表达水平。随后还进行了双荧光素酶报告基因检测。
与原代人肝细胞（hHC）相比，HuH7 细胞中 miR-31 的表达水平显著上调。双荧光素酶报告基因检测表明，IRF-1 是 miR-31 的直接靶标。IRF-1 的表达水平在 HuH7 和 HepG2 细胞系中降低。IRF-1 在 HCC 组织及其配对的相邻组织中与 miR-31 呈负相关。miR-31 的表达水平与 IRF-1 呈负相关。miR-31 抑制剂上调 HuH7 细胞中 IRF-1 的表达水平，而 miR-31 模拟物下调 HuH7 细胞中 IRF-1 的表达水平。此外，miR-31 模拟物抑制 IRF-1-3'UTR 报告基因活性，而 miR-31 抑制剂增强 IRF-1-3'UTR 报告基因活性，这取决于 miR-31 模拟物和 miR-31 抑制剂的浓度。
这些结果表明，miR-31 可以调节 HCC 中 IRF-1 的表达水平，这可能为靶向 miR-31 治疗 HCC 提供新的理论依据。
