Department of Gynecology, Jining No.1 People's Hospital, Jining 272000, Shandong, China.
Department of Hepatobiliary Surgery, Jining No.1 People's Hospital, Jining 272000, Shandong, China.
Biosci Rep. 2018 Jun 21;38(3). doi: 10.1042/BSR20180668. Print 2018 Jun 29.
Our previous study suggested that minichromosome maintenance protein 5 (MCM5) overexpression was observed in cervical adenocarcinoma and closely associated with advanced clinical stage, more metastatic lymph nodes, present distant metastasis, low histological grade, and poor prognosis. Down-regulation of MCM5 inhibited cervical adenocarcinoma cell proliferation. The purpose of the present study is to search and confirm valuable microRNAs (miRNAs), which target MCM5 to modulate cervical adenocarcinoma cell proliferation. In our results, we found that levels of miR-362-3p expression were reduced in cervical adenocarcinoma tissues and cell lines. Moreover, 3'-UTR of MCM5 had binding site of miR-362-3p through analyzing Targetscan database and miRanda database, and there were an inverse association between miR-362-3p and MCM5 in cervical adenocarcinoma tissues. Furthermore, we verified miR-362-3p directly targeted to 3'-UTR of DCLK1 by luciferase reporter assay, and negatively regulated mRNA and protein expressions of MCM5 by qPCR and Western blot. Then, we conducted gain-of-function study and rescued-function study, and found that miR-362-3p served as a tumor suppressive miRNA to modulate cervical adenocarcinoma cell proliferation through regulating the functional target MCM5. Finally, we analyzed correlations between miR-362-3p expression and clinicopathological characteristics and observed that miR-362-3p low expression was associated with advanced clinical stage and poor prognosis. In conclusion, miR-362-3p is a tumor suppressive miRNA in cervical adenocarcinoma.
我们之前的研究表明，微小染色体维持蛋白 5（MCM5）在宫颈腺癌中过表达，并且与晚期临床分期、更多转移性淋巴结、存在远处转移、低组织学分级和不良预后密切相关。MCM5 的下调抑制了宫颈腺癌细胞的增殖。本研究旨在寻找并确认有价值的 microRNAs（miRNAs），这些 miRNAs 以 MCM5 为靶点调节宫颈腺癌细胞的增殖。在我们的研究结果中，我们发现 miR-362-3p 在宫颈腺癌组织和细胞系中的表达水平降低。此外，通过分析 Targetscan 数据库和 miRanda 数据库，我们发现 MCM5 的 3'-UTR 具有 miR-362-3p 的结合位点，并且 miR-362-3p 与宫颈腺癌组织中的 MCM5 呈负相关。此外，我们通过荧光素酶报告基因实验验证了 miR-362-3p 直接靶向 DCLK1 的 3'-UTR，并通过 qPCR 和 Western blot 验证了 miR-362-3p 负调控 MCM5 的 mRNA 和蛋白表达。然后，我们进行了功能获得和功能恢复研究，发现 miR-362-3p 通过调节功能性靶标 MCM5 作为肿瘤抑制 miRNA 来调节宫颈腺癌细胞的增殖。最后，我们分析了 miR-362-3p 表达与临床病理特征之间的相关性，并观察到 miR-362-3p 低表达与晚期临床分期和不良预后相关。总之，miR-362-3p 是宫颈腺癌中的一种肿瘤抑制 miRNA。
