MP66-17 MICRORNA-224 IS DOWN-REGULATED IN PROSTATE CANCER AND MEDIATES TUMOR-SUPPRESSIVE EFFECTS IN VITRO

Abstract

You have accessJournal of UrologyProstate Cancer: Basic Research V1 Apr 2015MP66-17 MICRORNA-224 IS DOWN-REGULATED IN PROSTATE CANCER AND MEDIATES TUMOR-SUPPRESSIVE EFFECTS IN VITRO Felix Bienert, Kati Erdmann, Susanne Fuessel, and Manfred P. Wirth Felix BienertFelix Bienert More articles by this author , Kati ErdmannKati Erdmann More articles by this author , Susanne FuesselSusanne Fuessel More articles by this author , and Manfred P. WirthManfred P. Wirth More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.2370AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The microRNA miR-224 is a putative regulator of alpha-methylacyl-CoA racemase (AMACR) which is known to be up-regulated in prostate cancer (PCa). Here, we analyzed the expression levels of miR-224 and its host gene gamma-aminobutyric acid A receptor / epsilon subunit (GABRE). The functional role of miR-224 was then investigated in vitro. METHODS Expression levels were analyzed by qPCR using 50 malignant (PCa) and matched non-malignant tissue samples from prostatectomy explants as well as 30 samples derived from patients with benign prostatic hyperplasia (BPH). AMACR expression data were used from a previous study on the same sample cohort. DU145, PC-3 and LNCaP PCa cells were transfected with 100 nM of miR-224 mimic or a control construct for 4 h. Protein and mRNA expression levels of AMACR as well as functional analyses (viability, proliferation, cell colony formation, apoptosis) were conducted 48 h after transfection. RESULTS The expression of miR-224 was significantly decreased in PCa tissue samples by 3.5- and 2.7-fold compared to matched non-malignant and BPH tissue samples, respectively. Concomitantly, GABRE mRNA expression was completely down-regulated compared to either control group. The down-regulation of miR-224 significantly correlated with the down-regulation of its host gene GABRE (Spearman correlation coefficient: 0.442) as well as with the previously determined up-regulation of its target gene AMACR (Spearman correlation coefficient: 0.384). In vitro, miR-224 led to slight decreases of AMACR mRNA by 30% and protein by 20% only in PC-3 cells. MiR-224 marginally influenced cellular viability and proliferation in DU145 and LNCaP cells. In contrast, miR-224-treated PC-3 cells displayed a decrease in viability by 17% and proliferation by 34% compared to control. Cell colony formation was inhibited in both DU145 and PC-3 cells by 23% and 42%, respectively, following miR-224 administration. Furthermore, miR-224 produced a doubling of apoptosis-mediated cell death rate in PC-3 cells, whereas apoptosis rate remained unaffected in DU145 and LNCaP cells. CONCLUSIONS Our findings indicate that the PCa-specific down-regulation of miR-224 correlates with the down-regulation of its host gene GABRE as well as with the up-regulation of its putative target gene AMACR. Depending on the cell line miR-224 could inhibit tumor cell viability and growth as well as induce apoptosis-mediated cell death which suggests a tumor-suppressive role for this microRNA in PCa. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e822 Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information Felix Bienert More articles by this author Kati Erdmann More articles by this author Susanne Fuessel More articles by this author Manfred P. Wirth More articles by this author Expand All Advertisement Advertisement PDF DownloadLoading ...