Department of Hepatobiliary and Pancreatic Surgery, Shenzhen Hospital, Peking University, Shenzhen, Guangdong 518036, P.R. China.
Mol Med Rep. 2021 May;23(5). doi: 10.3892/mmr.2021.11957. Epub 2021 Mar 24.
Increasing evidence suggests that long noncoding RNAs (lncRNAs) influence the pathogenesis and progression of hepatocellular carcinoma (HCC). The authors of the present study previously reported that abnormal upregulation of lncRNA DQ786243 (lncDQ) was associated with poor prognoses for patients with HCC. However, the elucidation of underlying mechanisms which influenced these results was not completed. Thus, the current study aimed to characterize the mechanisms and functions of lncDQ that facilitate its promotion of HCC progression. lncDQ, miR‑15b‑5p and Wnt3A expression levels were characterized in HCC and portal vein tumor thrombus tissue samples and for liver cancer and liver cancer cell lines using reverse transcription‑quantitative PCR. Bioinformatics software was used for the analysis of interactions between lncDQ and miR‑15b‑5p, miR‑15b‑5p and Wnt3A. Luciferase assays confirmed the binding relationships between miR‑15b‑5p and the 3' untranslated region (UTR) of Wnt3A. Using online databases, prognostic values of miR‑15b‑5p and Wnt3A were also assessed. Proliferation and invasion assays were used to assess liver cancer and HCC cell functions after individually silencing lncDQ and miR‑15b‑5p expression in the cells. Western blotting was used for the investigation of alterations of the expression of Wnt3A/β‑catenin and epithelial‑mesenchymal transition (EMT) signal pathways. lncDQ and Wnt3A expression were significantly increased in HCC tissues, whereas miR‑15b‑5p was downregulated in HCC tissues. Low expression of miR‑15b‑5p was also associated with poor prognoses for patients with HCC. lncDQ was able to bind with miR‑15b‑5p and served as a competing endogenous RNA. As the target gene of miR‑15b‑5p, Wnt3A was correlated with poor prognoses for patients with HCC. Silencing of lncDQ expression significantly attenuated proliferation and invasion of liver cancer and HCC cells, however the inhibition of miR‑15b‑5p was able to reverse this effect. However, silencing of lncDQ and miR‑15b‑5p expression simultaneously resulted in the partial rescue of the inhibitory effect in the liver cancer and HCC cells. lncDQ inhibited miR‑15b‑5p so as to promote HCC cell invasion and proliferation through activation of the Wnt3A/β‑catenin/EMT pathway. Taken together, the results of the present study suggested that the lncDQ/miR‑15b‑5p axis modulates the progression of HCC.
越来越多的证据表明，长非编码 RNA（lncRNA）影响肝细胞癌（HCC）的发病机制和进展。本研究的作者先前报道，lncRNA DQ786243（lncDQ）的异常上调与 HCC 患者的预后不良有关。然而，影响这些结果的潜在机制尚未阐明。因此，本研究旨在描述促进 HCC 进展的 lncDQ 的作用机制和功能。采用逆转录-定量 PCR 检测 HCC 及门静脉癌栓组织样本及肝癌和肝癌细胞系中 lncDQ、miR-15b-5p 和 Wnt3A 的表达水平。采用生物信息学软件分析 lncDQ 与 miR-15b-5p 、miR-15b-5p 与 Wnt3A 之间的相互作用。荧光素酶报告基因实验证实 miR-15b-5p 与 Wnt3A 3'非翻译区（UTR）之间的结合关系。利用在线数据库评估 miR-15b-5p 和 Wnt3A 的预后价值。分别沉默细胞中的 lncDQ 和 miR-15b-5p 表达后，采用增殖和侵袭实验评估肝癌和 HCC 细胞功能。采用 Western blot 检测 Wnt3A/β-catenin 和上皮-间充质转化（EMT）信号通路表达的变化。结果显示，HCC 组织中 lncDQ 和 Wnt3A 表达水平升高，而 miR-15b-5p 表达水平降低。miR-15b-5p 低表达与 HCC 患者预后不良相关。lncDQ 可与 miR-15b-5p 结合并作为竞争内源性 RNA。作为 miR-15b-5p 的靶基因，Wnt3A 与 HCC 患者的预后不良相关。沉默 lncDQ 表达可显著抑制肝癌和 HCC 细胞的增殖和侵袭，但抑制 miR-15b-5p 可逆转这一作用。然而，同时沉默 lncDQ 和 miR-15b-5p 表达可部分挽救肝癌和 HCC 细胞的抑制作用。lncDQ 通过激活 Wnt3A/β-catenin/EMT 通路抑制 miR-15b-5p ，从而促进 HCC 细胞侵袭和增殖。综上所述，本研究结果表明，lncDQ/miR-15b-5p 轴调节 HCC 的进展。
