Yuan Renshun, Zhi Qiaoming, Zhao Hong, Han Ye, Gao Ling, Wang Bin, Kou Zhongyang, Guo Zhaoji, He Songbing, Xue Xiaofeng, Hu Hao
Department of General Surgery, The First Affiliated Hospital of Soochow University, 188# Shizi Road, Suzhou, 215006, China.
Tumour Biol. 2015 Apr;36(4):3093-100. doi: 10.1007/s13277-014-2945-2. Epub 2014 Dec 16.
Aberrant microRNA (miRNA) expression has been widely recognized to play an extremely important role in several cancers, including hepatocellular carcinoma (HCC). According to the previous studies, abnormal miR-106a expression was closely related to various cancer occurrences. However, the miR-106a expression in HCC remains unclear. In our study, we firstly detected the miR-106a expression levels in 36 pairs of HCC tissues. The results showed that miR-106a expression in HCC tissues was apparently higher than the level in the adjacent tissues. Then, we used quantitative real-time PCR (qPCR) and BSP to analyze miR-106a expression and promoter methylation in HCC cell lines. There came to a conclusion that the methylation status of the miR-106a promoter region was inversely correlated with the expression of miR-106a. After prediction with online software, we further used dual-luciferase reporter gene assay to ensure that TP53INP1 and CDKN1A might be the direct targets of miR-106a. At last, we explored the functions of miR-106a in HCC cells in vitro. Our results manifested that high-miR-106a cell line had stronger invasiveness, faster cell cycle progression, and more resistance to apoptosis compared with the low-miR-106a cell line. Therefore, our study suggested that upregulated expression of miR-106a by its promoter hypomethylation might contribute to the progression of HCC, which might be considered as a potentially effective biomarker and therapeutic approach in the future.
异常的微小RNA(miRNA)表达在包括肝细胞癌(HCC)在内的多种癌症中发挥着极其重要的作用,这一点已得到广泛认可。根据以往的研究,miR-106a的异常表达与各种癌症的发生密切相关。然而,HCC中miR-106a的表达情况仍不清楚。在我们的研究中,我们首先检测了36对HCC组织中miR-106a的表达水平。结果显示,HCC组织中miR-106a的表达明显高于相邻组织中的水平。然后,我们使用定量实时聚合酶链反应(qPCR)和亚硫酸氢盐测序法(BSP)分析HCC细胞系中miR-106a的表达和启动子甲基化情况。得出的结论是,miR-106a启动子区域的甲基化状态与miR-106a的表达呈负相关。通过在线软件预测后,我们进一步使用双荧光素酶报告基因检测来确定TP53INP1和CDKN1A可能是miR-106a的直接靶点。最后,我们在体外探索了miR-106a在HCC细胞中的功能。我们的结果表明,与低miR-106a细胞系相比,高miR-106a细胞系具有更强的侵袭性、更快的细胞周期进程以及对凋亡的更强抗性。因此,我们的研究表明,miR-106a启动子低甲基化导致的表达上调可能促进HCC的进展,这在未来可能被视为一种潜在有效的生物标志物和治疗方法。