子宫内膜癌中MMP16和TIMP2表达的转录后调控：miR-382、miR-410和miR-200b
Post-transcriptional Regulation of MMP16 and TIMP2 Expression miR-382, miR-410 and miR-200b in Endometrial Cancer.
作者信息
Rak Beata, Mehlich Dawid, Garbicz Filip, Domosud Zofia, Paskal Wiktor, Marczewska Janina M, Włodarski Paweł K
机构信息
Department of Histology and Embryology, Laboratory of Centre for Preclinical Research, Medical University of Warsaw, Warsaw, Poland.
Postgraduate School of Molecular Medicine, Warsaw, Poland.
出版信息
Cancer Genomics Proteomics. 2017 Sep-Oct;14(5):389-401. doi: 10.21873/cgp.20049.
BACKGROUND/AIM: The post-transcriptional regulation of matrix metalloproteinases (MMPs) via microRNAs (miRNAs) has been recently described in numerous human malignancies. However, the exact mechanisms of miRNA-mediated MMPs deregulation in endometrial cancer (EC) remain unclear. Herein, we aimed to analyze the expression of MMP2, MMP16 and TIMP2 and identify miRNAs that modulate their expression.
MATERIALS AND METHODS
Protein expression was assessed by immunohistochemistry in formalin-fixed paraffin-embedded EC samples. Target prediction algorithms were applied to select miRNAs binding the 3'UTRs of MMP16 (miR-377, miR-382, miR-410, miR-200b) or TIMP2 (miR-200b), and their levels were measured by qPCR in laser capture-microdissected tissue fragments. Luciferase assays and western blotting were used to indicate individual miRNA- mRNA interactions.
RESULTS
Overexpression of MMP2 and MMP16 in cancerous tissues corresponded to down-regulation of miR-377, miR-382 and miR-410, while decreased expression of TIMP2 was associated with miR-200b up-regulation. In vitro experiments confirmed direct regulation of MMP16 by miR-382 and miR-410, and TIMP2 by miR-200b in EC Ishikawa cells.
CONCLUSION
We demonstrated novel mechanisms of miRNA-mediated regulation of MMPs activity in EC.
背景/目的：近年来，在多种人类恶性肿瘤中已发现通过微小RNA（miRNA）对基质金属蛋白酶（MMP）进行转录后调控。然而，miRNA介导的子宫内膜癌（EC）中MMP失调的确切机制仍不清楚。在此，我们旨在分析MMP2、MMP16和TIMP2的表达，并鉴定调节它们表达的miRNA。
材料与方法
通过免疫组织化学对福尔马林固定石蜡包埋的EC样本中的蛋白质表达进行评估。应用靶标预测算法选择与MMP16（miR-377、miR-382、miR-410、miR-200b）或TIMP2（miR-200b）的3'非翻译区结合的miRNA，并通过qPCR在激光捕获显微切割的组织片段中测量它们的水平。荧光素酶测定和蛋白质印迹用于表明单个miRNA与mRNA的相互作用。
结果
癌组织中MMP2和MMP16的过表达与miR-377、miR-382和miR-410的下调相对应，而TIMP2表达的降低与miR-200b的上调相关。体外实验证实miR-382和miR-410在EC Ishikawa细胞中直接调节MMP16，miR-200b直接调节TIMP2。
结论
我们证明了miRNA介导的EC中MMP活性调节的新机制。
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