本研究旨在研究长链非编码RNA在肝细胞癌(HCC)转移性三阴性乳腺癌淋巴结(lncRNAHUMT)中高度上调的作用和机制。定量实时聚合酶链反应用于评估HUMT的表达,microRNA(miR)-455-5p,和低密度脂蛋白受体相关蛋白4(LRP4)在肝癌组织中的表达。进行集落形成和5-乙炔基-2'-脱氧尿苷测定以评估细胞增殖。进行Transwell测定以测量细胞迁移和侵袭。使用流式细胞术评估细胞周期分布。LRP4、增殖细胞核抗原(PCNA)、基质金属肽酶2(MMP-2),使用蛋白质印迹检测MMP-9。荧光素酶报告基因测定和RNA免疫沉淀测定用于确认miR-455-5p与HUMT或LRP4之间的靶标关联。在我们的研究中,肝癌组织和细胞中HUMT水平增强。细胞增殖,入侵,肝癌细胞的迁移受到HUMT敲低的抑制,HUMT的敲除将细胞阻滞在G1期,并降低PCNA的水平,MMP-2和MMP-9。MiR-455-5p是HUMT的靶标。miR-455-5p的低表达逆转了HUMT沉默对HCC细胞的抑制作用。LRP4是miR-455-5p的靶标,并被miR-455-5p负调控。此外,LRP4表达被HUMT正修饰,LRP4抑制HUMT沉默对肝癌细胞的抑制作用。总之,肝癌细胞增殖,入侵,移民受到HUMT击倒的限制,与miR-455-5p/LRP4轴相关。
The present study aimed at investigating the effects and mechanism of long noncoding RNA highly upregulated in metastatic triple-negative breast cancer lymph node (lncRNA HUMT) in hepatocellular carcinoma (HCC). Quantitative real-time polymerase chain reaction was used to assess the expression of HUMT, microRNA (miR)-455-5p, and low-density lipoprotein receptor-related protein 4 (
LRP4) in HCC tissues. Colony forming and 5-ethynyl-2\'-deoxyuridine assays were performed to assess cell proliferation. Transwell assay was performed to measure cell migration and invasion. Cell cycle distribution was assessed using flow cytometry. The protein expression of
LRP4, proliferating cell nuclear antigen (PCNA), matrix metallopeptidase 2 (MMP-2), and MMP-9 was detected using western blot. Luciferase reporter assay and RNA immunoprecipitation assay was used to confirm the target association between miR-455-5p and HUMT or
LRP4. In our study, the level of HUMT was enhanced in HCC tissues and cells. Cell proliferation, invasion, and migration in HCC cells were repressed by knockdown of HUMT, and knockdown of HUMT arrested cells in G1 phase and decreased the levels of PCNA, MMP-2, and MMP-9. MiR-455-5p was a target of HUMT. Lowexpression of miR-455-5p reversed the inhibitive influence on HCC cells induced by of HUMT silencing.
LRP4 was a target of miR-455-5p and was negatively regulated by miR-455-5p. In addition,
LRP4 expression was positively modified by HUMT, and
LRP4 inhibited the inhibitory effects on HCC cells induced by HUMT silencing. In conclusion, HCC cell proliferation, invasion, and migration were restrained by knockdown of HUMT, which was related to the miR-455-5p/LRP4 axis.