Ding Gangqiang, Peng Zhen, Shang Jia, Kang Yi, Ning Huibin, Mao Chongshan
Department of Infectious Diseases, People's Hospital of Zhengzhou University, Henan Provincial People's Hospital, Zhengzhou, China.
Onco Targets Ther. 2017 Jun 30;10:3241-3247. doi: 10.2147/OTT.S134910. eCollection 2017.
In the previous study, it was found that long intergenic noncoding RNA-p21 (lincRNA-p21) was downregulated in hepatocellular carcinoma (HCC) and lincRNA-p21 overexpression inhibited tumor invasion through inducing epithelial-mesenchymal transition. However, the underlying mechanism was not fully elaborated. In this study, lincRNA-p21 expression was measured in 12 paired HCC and nontumor adjacent normal tissues by quantitative real-time polymerase chain reaction. The effects of lincRNA-p21 on HCC cells were studied using lentivirus expressing lincRNA-p21 vector in vitro. The association between lincRNA-p21 level and miR-9 level was tested with the Spearman rank correlation. The effects of miR-9 on HCC cells were studied by using miR-9 inhibitor in vitro. Luciferase assay was used to validate the target of miR-9. The results showed that lincRNA-p21 was downregulated in human HCC tissues and cell lines. LincRNA-p21 overexpression significantly inhibited HCC cell migration and invasion in vitro. Besides, lincRNA-p21 negatively regulated miR-9 expression level, and miR-9 was upregulated in human HCC tissues and cells. MiR-9 knockdown inhibited HCC cell migration and invasion in vitro. Finally, the luciferase assay results showed that E-cadherin was a direct target of miR-9. The expression level of E-cadherin was found to be regulated by lincRNA-p21 and miR-9. Altogether, the results suggested that lincRNA-p21 inhibits migration and invasion of HCC cells through regulating miR-9-mediated E-cadherin cascade signaling pathway.
在先前的研究中，发现长链基因间非编码RNA-p21（lincRNA-p21）在肝细胞癌（HCC）中表达下调，lincRNA-p21过表达通过诱导上皮-间质转化抑制肿瘤侵袭。然而，其潜在机制尚未完全阐明。在本研究中，通过定量实时聚合酶链反应检测了12对HCC组织及癌旁非肿瘤正常组织中lincRNA-p21的表达。体外使用表达lincRNA-p21载体的慢病毒研究lincRNA-p21对HCC细胞的影响。采用Spearman等级相关检验lincRNA-p21水平与miR-9水平之间的关联。体外使用miR-9抑制剂研究miR-9对HCC细胞的影响。采用荧光素酶报告基因检测法验证miR-9的靶标。结果显示，lincRNA-p21在人HCC组织和细胞系中表达下调。lincRNA-p21过表达显著抑制体外HCC细胞的迁移和侵袭。此外，lincRNA-p21负向调节miR-9的表达水平，且miR-9在人HCC组织和细胞中上调。敲低miR-9可抑制体外HCC细胞的迁移和侵袭。最后，荧光素酶报告基因检测结果显示E-钙黏蛋白是miR-9的直接靶标。发现E-钙黏蛋白的表达水平受lincRNA-p21和miR-9的调控。总之，结果表明lincRNA-p21通过调控miR-9介导的E-钙黏蛋白级联信号通路抑制HCC细胞的迁移和侵袭。
