Department of Pathology, Medical College of Yan'an University, Yan'an, Shanxi 716000, P.R. China.
Department of Pharmacology, Medical College of Yan'an University, Yan'an, Shanxi 716000, P.R. China.
Mol Med Rep. 2019 Apr;19(4):2913-2920. doi: 10.3892/mmr.2019.9934. Epub 2019 Feb 5.
Previous studies have demonstrated that numerous tumor‑specific microRNAs (miRNAs) are upregulated or downregulated in hepatocellular carcinoma (HCC), and that their dysregulation is implicated in HCC occurrence and development. Therefore, investigation of crucial miRNAs involved in HCC oncogenesis and progression may provide novel insights into the therapy of patients with this malignant tumor. In the present study, reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) assays were performed to detect tissue and cellular expression levels of miRNA‑663b (miR‑663b) in HCC. The effects of miR‑663b overexpression on the proliferation and invasion of HCC cells were examined using Cell Counting Kit‑8 and Transwell invasion assays, respectively. The direct target of miR‑663b in HCC cells was determined by bioinformatics analysis, luciferase reporter assay, RT‑qPCR and western blot analysis. It was observed that miR‑663b was expressed at low levels in HCC tissues and cell lines. miR‑663b upregulation suppressed the proliferative and invasive abilities of HCC cells. Additionally, Grb2‑associated binding 2 (GAB2) was regarded as a direct target gene of miR‑663b in HCC cells. Furthermore, GAB2 was overexpressed in HCC tissues, and overexpression of GAB2 was inversely correlated with levels of miR‑663b. GAB2 overexpression was able to rescue the suppressive effects of miR‑663b on HCC cells. These results demonstrated that this newly‑identified miR‑663b/GAB2 axis may be implicated in HCC occurrence and development.
先前的研究表明，许多肿瘤特异性 microRNAs（miRNAs）在肝细胞癌（HCC）中上调或下调，其失调与 HCC 的发生和发展有关。因此，研究与 HCC 致癌和进展相关的关键 miRNAs 可能为治疗这种恶性肿瘤的患者提供新的见解。在本研究中，通过逆转录-定量聚合酶链反应（RT-qPCR）检测 HCC 组织和细胞中 miRNA-663b（miR-663b）的表达水平。通过细胞计数试剂盒-8 和 Transwell 侵袭试验分别检测 miR-663b 过表达对 HCC 细胞增殖和侵袭的影响。通过生物信息学分析、荧光素酶报告基因检测、RT-qPCR 和 Western blot 分析确定 HCC 细胞中 miR-663b 的直接靶基因。结果观察到 miR-663b 在 HCC 组织和细胞系中表达水平较低。miR-663b 的上调抑制了 HCC 细胞的增殖和侵袭能力。此外，Grb2 相关结合蛋白 2（GAB2）被认为是 HCC 细胞中 miR-663b 的直接靶基因。此外，GAB2 在 HCC 组织中过度表达，并且 GAB2 的过表达与 miR-663b 的水平呈负相关。GAB2 的过表达能够挽救 miR-663b 对 HCC 细胞的抑制作用。这些结果表明，这个新发现的 miR-663b/GAB2 轴可能与 HCC 的发生和发展有关。
