title: Reprimo as a modulator of cell migration and invasion in the MDA-MB-231 breast cancer cell line.
abstract: BACKGROUND
  Reprimo (RPRM), a highly glycosylated protein, is a new downstream effector of p53-induced cell cycle arrest at the G2/M checkpoint, and a putative tumor suppressor gene frequently silenced via methylation of its promoter region in several malignances. The aim of this study was to characterize the epigenetic inactivation and its biological function in BC cell lines.
  
  
  METHODS
  The correlation between RPRM methylation and loss of mRNA expression was assessed in six breast cancer cell lines by methylation specific PCR (MSP), 5'-Aza-2'-deoxycytidine treatment and RT-PCR assays. MDA-MB-231 cells were chosen to investigate the phenotypic effect of RPRM in cell proliferation, cell cycle, cell death, cell migration and invasion.
  
  
  RESULTS
  In the cancer methylome system (CMS) (web-based system for visualizing and analyzing genome-wide methylation data of human cancers), the CpG island region of RPRM (1.1 kb) was hypermethylated in breast cancer compared to normal breast tissue; more interesting still was that ERα(+) tumors showed higher methylation intensity than ERα(-). Downregulation of RPRM mRNA by methylation was confirmed in MDA-MB-231 and BT-20 cell lines. In addition, overexpression of RPRM in MDA-MB-231 cells resulted in decreased rates of cell migration, wound healing and invasion in vitro. However, RPRM overexpression did not alter cell viability, phosphatidylserine (PS) translocation or G2/M cell cycle transition.
  
  
  CONCLUSIONS
  Taken together, these data suggest that RPRM is involved in decreased cell migration and invasion in vitro, acting as a potential tumor suppressor gene in the MDA-MB-231 cell line.
authors.affiliation: Department of Pathology, Molecular Pathology Laboratory BIOREN-CEGIN, School of Medicine, Universidad de La Frontera, Temuco, Chile. k.buchegger@gmail.com.
authors.forename: Kurt
authors.initials: K
authors.lastname: Buchegger
authors.affiliation: Department of Pathology, Molecular Pathology Laboratory BIOREN-CEGIN, School of Medicine, Universidad de La Frontera, Temuco, Chile. carmengloriaili@gmail.com.
authors.forename: Carmen
authors.initials: C
authors.lastname: Ili
authors.affiliation: Department of Pathology, Molecular Pathology Laboratory BIOREN-CEGIN, School of Medicine, Universidad de La Frontera, Temuco, Chile. 83ismode@gmail.com.
authors.forename: Ismael
authors.initials: I
authors.lastname: Riquelme
authors.affiliation: School of Health Sciences, Universidad Católica de Temuco, Temuco, Chile. pablolete@gmail.com.
authors.forename: Pablo
authors.initials: P
authors.lastname: Letelier
authors.affiliation: Centre for Translational Research in Oncology (CITO) and Department of Hematology and Oncology, Pontificia Universidad Catolica de Chile, Santiago, Chile. corvalan01@gmail.com.
authors.forename: Alejandro H
authors.initials: AH
authors.lastname: Corvalán
authors.affiliation: Department of Pathology, Molecular Pathology Laboratory BIOREN-CEGIN, School of Medicine, Universidad de La Frontera, Temuco, Chile. brebimieville@gmail.com.
authors.forename: Priscilla
authors.initials: P
authors.lastname: Brebi
authors.affiliation: Department of Molecular Medicine/Institute of Biotechnology, University of Texas Health Science Center at San Antonio, STRF, Room 225, 7703 Floyd Curl Drive, San Antonio, TX, 78229, USA. huangt3@uthscsa.edu.
authors.forename: Tim Hui-Ming
authors.initials: TH
authors.lastname: Huang
authors.affiliation: Department of Pathology, Advanced Center for Chronic Diseases (ACCDiS) (CITO), School of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile. jcroas@gmail.com.
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Roa
mesh_terms: D000704:Analysis of Variance; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000235:genetics / Q000473:pathology*; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000235:genetics / Q000502:physiology*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics / Q000502:physiology*; D049109:Cell Proliferation / Q000235:genetics / Q000502:physiology*; D002470:Cell Survival; D019175:DNA Methylation; D044127:Epigenesis, Genetic; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006023:Glycoproteins / Q000235:genetics / Q000502:physiology*; D006801:Humans; D009361:Neoplasm Invasiveness; D060888:Real-Time Polymerase Chain Reaction; D018709:Statistics, Nonparametric
chemical_list: D018797:Cell Cycle Proteins; D006023:Glycoproteins; C412566:RPRM protein, human