Wang Yuping, Hu Yuna, Guo Junhui, Wang Lingling
First Oncology Ward, Henan Province Hospital of TCM, Zhengzhou, P.R. China.
Genet Test Mol Biomarkers. 2019 Feb;23(2):98-104. doi: 10.1089/gtmb.2018.0285.
To identify whether miR-148a-3p interacts with DNA (cytosine-5)-methyltransferase 1 (DNMT1) in esophageal cancer.
A luciferase assay and immunoblotting were performed to detect the relationship between miR-148a-3p and DNMT1. The MTT method, Annexin V/propidium iodide staining, and Transwell assays were adopted to assess the biological behaviors in EC109 cells. The association between the expression level of miR-148a-3p, clinical features, and prognosis were evaluated by chi-square test and univariate survival analysis.
In this study, DNMT1 was identified as a direct target of miR-148a-3p by luciferase assay and Western blot. Real-time quantitative PCR analyses showed that the relative expression levels of miR-148a-3p and DNMT1 were reduced in esophageal cancer samples compared with adjacent tissues; and a negative relationship between both was indicated. Upon overexpression of miR-148a-3p in esophageal cancer cells, proliferation and invasion were significantly suppressed, and apoptosis was promoted. A higher level of miR-148a-3p was correlated with better patient outcomes.
Our study indicated that miR-148a-3p, by targeting DNMT1, likely regulates cell proliferation and invasion in esophageal cancer. miR-148a-3p might also be used prognostically in esophageal cancer and serve as a therapeutic target in the future.
确定在食管癌中miR-148a-3p是否与DNA(胞嘧啶-5)-甲基转移酶1(DNMT1)相互作用。
进行荧光素酶报告基因检测和免疫印迹以检测miR-148a-3p与DNMT1之间的关系。采用MTT法、膜联蛋白V/碘化丙啶染色和Transwell实验评估EC109细胞的生物学行为。通过卡方检验和单因素生存分析评估miR-148a-3p表达水平与临床特征及预后之间的关联。
在本研究中,通过荧光素酶报告基因检测和蛋白质免疫印迹确定DNMT1是miR-148a-3p的直接靶点。实时定量PCR分析显示,与癌旁组织相比,食管癌样本中miR-148a-3p和DNMT1的相对表达水平降低;且二者呈负相关。在食管癌细胞中过表达miR-148a-3p后,细胞增殖和侵袭受到显著抑制,细胞凋亡增加。miR-148a-3p水平较高与患者较好的预后相关。
我们的研究表明,miR-148a-3p通过靶向DNMT1可能调控食管癌细胞的增殖和侵袭。miR-148a-3p也可能在食管癌预后评估中发挥作用,并有望成为未来的治疗靶点。