Department of Pediatric Surgery, The Affiliated Hospital of Medical College Qingdao University, Qingdao, China.
FEBS J. 2013 Feb;280(3):855-66. doi: 10.1111/febs.12074. Epub 2013 Jan 16.
In this study, we found that the expression of miR-15a was positively correlated with neuroblastoma (NB) clinical pathological stage and was negatively correlated with reversion-inducing cysteine-rich protein with Kazal motifs (RECK) expression. Using the enhanced green fluorescent protein (EGFP) reporter construct carrying the 3'-UTR of RECK, we identified RECK as a direct target of miR-15a. Suppression of miR-15a significantly decreased the migration ability of GI-LA-N and SK-N-SH cell lines, whereas overexpression of miR-15a increased the migration ability; these effects could be partly reversed by RECK inhibition or ectopic expression. Moreover, inhibition of miR-15a significantly increased secreted matrix metalloproteinase-9 expression in culture medium through regulating the expression of RECK. These findings provide new insights into the characteristics of the miR-15a-RECK-matrix metalloproteinase-9 axis in NB progression, especially in NB migration and invasion.
在这项研究中,我们发现 miR-15a 的表达与神经母细胞瘤(NB)的临床病理分期呈正相关,与富含半胱氨酸的逆转诱导蛋白与 Kazal 基序(RECK)的表达呈负相关。利用携带 RECK 3'-UTR 的增强型绿色荧光蛋白(EGFP)报告构建体,我们确定 RECK 是 miR-15a 的直接靶标。抑制 miR-15a 可显著降低 GI-LA-N 和 SK-N-SH 细胞系的迁移能力,而过表达 miR-15a 则增加了迁移能力;RECK 的抑制或异位表达部分逆转了这些效应。此外,通过调节 RECK 的表达,抑制 miR-15a 可显著增加培养上清液中分泌型基质金属蛋白酶-9 的表达。这些发现为 NB 进展中 miR-15a-RECK-基质金属蛋白酶-9 轴的特征提供了新的见解,尤其是在 NB 的迁移和侵袭中。