title: Activin A regulates microRNAs and gene expression in LNCaP cells.
abstract: Prostate cancer (PCa) is an increasing health issue worldwide. For patients with advanced castration-resistant PCa (CRPC) treatment options are limited and overall survival is relatively short. Paired with this, non-invasive diagnostic options are yet to be established. Activins are members of the TGF-β superfamily and have been linked to prostate physiology. For instance, activin A is an inhibitor of growth in the prostate. A novel class of non-coding RNA, microRNAs (miRNAs) have been intrinsically linked to a range of cellular processes and carcinogenesis. No studies have investigated the impact of activin A on miRNA expression in PCa cell lines. Hence, the objective of this study was to determine the effect of activin A on miRNA expression and downstream target genes in PCa.
  
  
  
  Activin-sensitive (LNCaP) and insensitive (PC3) prostate cells were treated with 50 ng/ml of activin A for 72 hr. To examine miRNA expression following treatment, SYBR RT-qPCR miRNA arrays were used in conjunction with TaqMan RT-qPCR. MiRPath-TarBase analysis was conducted using the miRNAs that were significantly altered following activin A treatment of LNCaP cells to highlight enriched target genes within biological pathways. Highlighted target genes were assessed using pathway-focused TGF-β and cell cycle SYBR RT-qPCR arrays.
  
  
  
  Activin A treatment altered nine miRNAs in LNCaP cells: miR-222-3p, miR-15b-5p, miR-93-5p, miR-18a-5p, and let-7i-5p were significantly decreased, while miR-30a/30d-5p, let-7c, and miR-196b-5p were significantly increased versus media control. In PC3 cells five miRNAs were altered: miR-130a-3p, miR-7-5p, and miR-140-3p were significantly decreased while miR-191-5p and miR-26a-5p were significantly increased versus media control. MiRPath-TarBase analysis highlighted that the miRNAs significantly altered in LNCaP cells targeted genes contained in activin A-related KEGG pathways. Furthermore, when LNCaP cells were treated with activin A the expression of the targeted genes was the inverse of the expression of activin A-mediated miRNAs.
  
  
  
  This study demonstrated the ability of activin A to modulate miRNA expression in PCa cell lines and suggests a correlative relationship between miRNA expression and downstream target genes in LNCaP cells. This study provides impetus for further studies into activin A and miRNAs in PCa. Prostate 76:951-963, 2016. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Department of Anatomy, University of Otago, Dunedin, New Zealand.
authors.forename: Edward Christopher
authors.initials: EC
authors.lastname: Ottley
authors.affiliation: Department of Anatomy, University of Otago, Dunedin, New Zealand.
authors.forename: Helen Diana
authors.initials: HD
authors.lastname: Nicholson
authors.affiliation: Department of Anatomy, University of Otago, Dunedin, New Zealand.
authors.forename: Elspeth Joan
authors.initials: EJ
authors.lastname: Gold
mesh_terms: D028341:Activins / Q000494:pharmacology*; D002453:Cell Cycle / Q000235:genetics; D045744:Cell Line, Tumor; D015870:Gene Expression / Q000187:drug effects*; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D019394:Ki-67 Antigen / Q000235:genetics; D008297:Male; D035683:MicroRNAs / Q000032:analysis / Q000235:genetics*; D016133:Polymerase Chain Reaction; D018809:Proliferating Cell Nuclear Antigen / Q000235:genetics; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000473:pathology; D016212:Transforming Growth Factor beta / Q000235:genetics
chemical_list: D019394:Ki-67 Antigen; D035683:MicroRNAs; D018809:Proliferating Cell Nuclear Antigen; D016212:Transforming Growth Factor beta; C422405:activin A; D028341:Activins
keywords: activin A; activins; cancer; microRNA; prostate cancer