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Roles of MicroRNA-21 and MicroRNA-29a in Regulating Cell Adhesion Related Genes in Bone Metastasis Secondary to Prostate Cancer

  • Mohamad, Maisarah (Department of Physiology, Faculty of Medicine, Hospital UKM) ;
  • Wahab, Norhazlina Abdul (Department of Physiology, Faculty of Medicine, Hospital UKM) ;
  • Yunus, Rosna (Histopathology Unit, Department of Pathology, Hospital Kuala Lumpur) ;
  • Murad, Nor AzianAbdul (UKM Molecular Biology Institute (UMBI), Hospital UKM) ;
  • Zainuddin, Zulkifli Md (Urology Unit, Department of Surgery, Universiti Kebangsaan Malaysia Medical Centre, Universiti Kebangsaan Malaysia (UKM)) ;
  • Sundaram, Murali (Urology Department, Hospital Kuala Lumpur) ;
  • Mokhtar, Norfilza Mohd (Department of Physiology, Faculty of Medicine, Hospital UKM)
  • Published : 2016.07.01

Abstract

Background: There is an increasing concern in the role of microRNA (miRNA) in the pathogenesis of bone metastasis (BM) secondary to prostate cancer (CaP). In this exploratory study, we hypothesized that the expression of vinculin (VCL) and chemokine X3C ligand 1 (CX3CL1) might be down-regulated in clinical samples, most likely due to the post-transcriptional modification by microRNAs. Targeted genes would be up-regulated upon transfection of the bone metastatic prostate cancer cell line, PC3, with specific microRNA inhibitors. Materials and Methods: MicroRNA software predicted that miR-21 targets VCL while miR-29a targets CX3CL1. Twenty benign prostatic hyperplasia (BPH) and 16 high grade CaP formalin-fixed paraffin embedded (FFPE) specimens were analysed. From the bone scan results, high grade CaP samples were further classified into CaP with no BM and CaP with BM. Transient transfection with respective microRNA inhibitors was done in both RWPE-1 (normal) and PC3 cell lines. QPCR was performed in all FFPE samples and transfected cell lines to measure VCL and CX3CL1 levels. Results: QPCR confirmed that VCL messenger RNA (mRNA) was significantly down-regulated while CX3CL1 was up-regulated in all FFPE specimens. Transient transfection with microRNA inhibitors in PC3 cells followed by qPCR of the targeted genes showed that VCL mRNA was significantly upregulated while CX3CL1 mRNA was significantly down-regulated compared to the RWPE-1 case. Conclusions: The down-regulation of VCL in FFPE specimens is most likely regulated by miR-21 based on the in vitro evidence but the exact mechanism of how miR-21 can regulate VCL is unclear. Up-regulated in CaP, CX3CL1 was found not regulated by miR-29a. More microRNA screening is required to understand the regulation of this chemokine in CaP with bone metastasis. Understanding miRNA-mRNA interactions may provide additional knowledge for individualized study of cancers.

Keywords

Prostate cancer;bone metastasis;miR-21;miR-29a;VCL;CX3CL1

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