Department of Medical Interventional Oncology, Yantai Qishan Hospital, Yantai, Shandong, China.
Department of Medical Gastroenterology, Yantai Qishan Hospital, Yantai, Shandong, China.
Bioengineered. 2022 Apr;13(4):8051-8063. doi: 10.1080/21655979.2022.2051841.
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.
本研究旨在探讨转移性三阴性乳腺癌淋巴结高表达的长非编码 RNA（lncRNA HUMT）在肝细胞癌（HCC）中的作用和机制。采用实时定量聚合酶链反应检测 HCC 组织中 HUMT、微小 RNA（miR）-455-5p 和低密度脂蛋白受体相关蛋白 4（LRP4）的表达。采用集落形成和 5-乙炔基-2'-脱氧尿苷实验评估细胞增殖。Transwell 实验用于检测细胞迁移和侵袭。采用流式细胞术评估细胞周期分布。采用 Western blot 检测 LRP4、增殖细胞核抗原（PCNA）、基质金属蛋白酶 2（MMP-2）和 MMP-9 的蛋白表达。采用荧光素酶报告基因实验和 RNA 免疫沉淀实验证实 miR-455-5p 与 HUMT 或 LRP4 之间的靶基因关联。在本研究中，HUMT 在 HCC 组织和细胞中上调。HUMT 敲低抑制 HCC 细胞的增殖、侵袭和迁移，HUMT 敲低使细胞停滞在 G1 期并降低 PCNA、MMP-2 和 MMP-9 的水平。miR-455-5p 是 HUMT 的靶基因。miR-455-5p 低表达逆转了 HUMT 沉默对 HCC 细胞的抑制作用。LRP4 是 miR-455-5p 的靶基因，受 miR-455-5p 负调控。此外，LRP4 表达受 HUMT 正向调节，LRP4 抑制了 HUMT 沉默对 HCC 细胞的抑制作用。综上所述，HUMT 敲低抑制 HCC 细胞的增殖、侵袭和迁移，与 miR-455-5p/LRP4 轴有关。
