Jiang Xinghua, Shen Xiaoxu
The affiliated hospital of Guizhou medical university, department of infectious diseases, Guiyang, 550004 Guizhou, China.
The affiliated hospital of Guizhou medical university, department of infectious diseases, Guiyang, 550004 Guizhou, China.
Bull Cancer. 2018 Oct;105(10):873-883. doi: 10.1016/j.bulcan.2018.07.013. Epub 2018 Sep 25.
Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide. MiR-299-5p has been demonstrated to play important roles in multiple human cancers. Nevertheless, little is known about the detailed function and molecular mechanism of miR-299-5p on HCC progression.
Quantitative real-time PCR (qRT-PCR) assay was used to assess the expression patterns of miR-299-5p and siah E3 ubiquitin protein ligase 1 (SIAH1) in HCC tissues and cell lines. Loss-of-function experiments were performed to explore the effect of miR-299-5p on HCC progression in vitro and in vivo. Target predicted by software algorithms, the connection between miR-299-5p and SIAH1 was verified by dual-luciferase reporter assay, qRT-PCR and western blot analysis. Subsequently, anti-miR-299-5p and si-SIAH1 were cotransfected into LM9 and Huh-7 cells to further explore whether the regulatory effect of miR-299-5p on HCC was mediated by SIAH1.
qRT-PCR assay revealed that miR-299-5p was upregulated and SIAH1 was downregulated in HCC tissues and cell lines. Moreover, miR-299-5p knockdown suppressed HCC progression in vitro and in vivo. In addition, anti-miR-299-5p-mediated regulatory effect on HCC cells was abated following the restoration of SIAH1 expression.
MiR-299-5p knockdown suppressed the progression of HCC by targeting SIAH1, highlighting its role as a potential biomarker and therapeutic target of HCC.
肝细胞癌(HCC)是全球最常见的恶性肿瘤之一。已证明miR-299-5p在多种人类癌症中发挥重要作用。然而,关于miR-299-5p对HCC进展的详细功能和分子机制知之甚少。
采用定量实时PCR(qRT-PCR)检测法评估miR-299-5p和SIAH E3泛素蛋白连接酶1(SIAH1)在HCC组织和细胞系中的表达模式。进行功能丧失实验以探讨miR-299-5p对体外和体内HCC进展的影响。通过软件算法预测靶点,采用双荧光素酶报告基因检测、qRT-PCR和蛋白质印迹分析验证miR-299-5p与SIAH1之间的联系。随后,将抗miR-299-5p和si-SIAH1共转染到LM9和Huh-7细胞中,以进一步探讨miR-299-5p对HCC的调节作用是否由SIAH1介导。
qRT-PCR检测显示,HCC组织和细胞系中miR-299-5p上调,SIAH1下调。此外,敲低miR-299-5p可抑制体外和体内HCC进展。此外,恢复SIAH1表达后,抗miR-299-5p介导的对HCC细胞的调节作用减弱。
敲低miR-299-5p通过靶向SIAH1抑制HCC进展,突出了其作为HCC潜在生物标志物和治疗靶点的作用。