title: TWIST1 upregulates the MAGEA4 oncogene.
abstract: Overexpression of MAGEA4 oncogene has been demonstrated in different malignancies; however, little is known about its exact mechanism for overexpression. TWIST1, as a bHLH transcription factor, activates a cell migration-invasion program involved in both embryonic and tumor development. Since MAGEA4 overexpression was statistically correlated to TWIST1, we aimed to elucidate the probable regulatory role of TWIST1 on MAGEA4 expression in KYSE30 cells.
  
  
  
  Expression pattern of MAGEA4 and TWIST1 was analyzed in 55 ESCC patients using relative comparative real-time PCR. In silico analysis of the MAGEA4 gene was performed. Methylation status of MAGEA4 promoter was determined by quantitative methylation specific PCR (qMSP). Using a retroviral system, KYSE30 cells were transduced to ectopically express TWIST1, followed by qRT-PCR, Western blot analysis, chromatin immunoprecipitation (ChIP), and luciferase assays to elucidate the regulatory role of TWIST1 on MAGEA4 gene expression.
  
  
  
  Concomitant overexpression of MAGEA4 and TWIST1 was detected in ESCC in significant correlation with each other in different clinicopathological indices of poor prognosis (P < 0.05). The TWIST1-expressing cells showed significantly higher MAGEA4 expression compared to control cells. ChIP and luciferase assays results confirmed indirect binding of TWIST1 to the E-boxes of MAGEA4 promoter sequence and revealed a novel regulatory role of TWIST1 in MAGEA4 upregulation.
  
  
  
  Since MAGEA4 is a highly expressed oncogene in a variety of malignancies in significant correlation with tumor cell invasiveness and aggressiveness, our finding may help understand one regulatory mechanism of increased expression in tumor cells. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Cellular and Molecular Research Center, Sabzevar University of Medical Sciences, Sabzevar, Iran.
authors.forename: Mohammad Mahdi
authors.initials: MM
authors.lastname: Forghanifard
authors.affiliation: Cellular and Molecular Research Center, Sabzevar University of Medical Sciences, Sabzevar, Iran.
authors.forename: Abolfazl
authors.initials: A
authors.lastname: Rad
authors.affiliation: Molecular Medicine Research Department, ACECR-Khorasan Razavi Branch, Mashhad, Iran.
authors.forename: Moein
authors.initials: M
authors.lastname: Farshchian
authors.affiliation: Division of Human Genetics, Immunology Research Center, Avicenna Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Maryam
authors.initials: M
authors.lastname: Khaleghizadeh
authors.affiliation: Division of Human Genetics, Immunology Research Center, Avicenna Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Mehran
authors.initials: M
authors.lastname: Gholamin
authors.affiliation: Division of Human Genetics, Immunology Research Center, Avicenna Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Meysam
authors.initials: M
authors.lastname: Moghbeli
authors.affiliation: Division of Human Genetics, Immunology Research Center, Avicenna Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Abbaszadegan
mesh_terms: D000951:Antigens, Neoplasm / Q000235:genetics* / Q000378:metabolism; D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D003198:Computer Simulation; D019175:DNA Methylation; D004938:Esophageal Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D000077277:Esophageal Squamous Cell Carcinoma; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D009363:Neoplasm Proteins / Q000235:genetics* / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism*; D011379:Prognosis; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D051879:Twist-Related Protein 1 / Q000235:genetics* / Q000378:metabolism*; D015854:Up-Regulation
chemical_list: D000951:Antigens, Neoplasm; C095204:MAGEA4 protein, human; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; C103173:TWIST1 protein, human; D051879:Twist-Related Protein 1
keywords: E-box; MAGEA4; TWIST1; cancer testis antigens; esophageal squamous cell carcinoma