title: Global DNA methylation profiling uncovers distinct methylation patterns of protocadherin alpha4 in metastatic and non-metastatic rhabdomyosarcoma.
abstract: Rhabdomyosarcoma (RMS), which can be classified as embryonal RMS (ERMS) and alveolar RMS (ARMS), represents the most frequent soft tissue sarcoma in the pediatric population; the latter shows greater aggressiveness and metastatic potential with respect to the former. Epigenetic alterations in cancer include DNA methylation changes and histone modifications that influence overall gene expression patterns. Different tumor subtypes are characterized by distinct methylation signatures that could facilitate early disease detection and greater prognostic accuracy.
  
  
  
  A genome-wide approach was used to examine methylation patterns associated with different prognoses, and DNA methylome analysis was carried out using the Agilent Human DNA Methylation platform. The results were validated using bisulfite sequencing and 5-aza-2'deoxycytidine treatment in RMS cell lines. Some in vitro functional studies were also performed to explore the involvement of a target gene in RMS tumor cells.
  
  
  
  In accordance with the Intergroup Rhabdomyosarcoma Study (IRS) grouping, study results showed that distinct methylation patterns distinguish RMS subgroups and that a cluster of protocadherin genes are hypermethylated in metastatic RMS. Among these, PCDHA4, whose expression was decreased by DNA methylation, emerged as a down-regulated gene in the metastatic samples. As PCDHA4-silenced cells have a significantly higher cell proliferation rate paralleled by higher cell invasiveness, PCDHA4 seems to behave as a tumor suppressor in metastatic RMS.
  
  
  
  Study results demonstrated that DNA methylation patterns distinguish between metastatic and non-metastatic RMS and suggest that epigenetic regulation of specific genes could represent a novel therapeutic target that could enhance the efficiency of RMS treatments.
authors.affiliation: Department of Biology, University of Padova, Padova, Italy. lucia.tombolan@unipd.it.|Department of Women's and Children's Health, University of Padova, Padova, Italy. lucia.tombolan@unipd.it.
authors.forename: L
authors.identifier: 0000-0002-8208-6244
authors.initials: L
authors.lastname: Tombolan
authors.affiliation: Department of Women's and Children's Health, University of Padova, Padova, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Poli
authors.affiliation: Department of Biology, University of Padova, Padova, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Martini
authors.affiliation: Institute of Pediatric Research, IRP, Padova, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Zin
authors.affiliation: C.R.I.B.I.-Biotechnology Centre, University of Padova, Padova, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Millino
authors.affiliation: C.R.I.B.I.-Biotechnology Centre, University of Padova, Padova, Italy.
authors.forename: B
authors.initials: B
authors.lastname: Pacchioni
authors.affiliation: C.R.I.B.I.-Biotechnology Centre, University of Padova, Padova, Italy.
authors.forename: B
authors.initials: B
authors.lastname: Celegato
authors.affiliation: Department of Women's and Children's Health, Hematology Oncology Division, University of Padova, Padova, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Bisogno
authors.affiliation: Department of Biology, University of Padova, Padova, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Romualdi
authors.affiliation: Department of Women's and Children's Health, Hematology Oncology Division, University of Padova, Padova, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Rosolen
authors.affiliation: Department of Biology, University of Padova, Padova, Italy. gerolamo.lanfranchi@unipd.it.|C.R.I.B.I.-Biotechnology Centre, University of Padova, Padova, Italy. gerolamo.lanfranchi@unipd.it.
authors.forename: G
authors.initials: G
authors.lastname: Lanfranchi
mesh_terms: D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D001706:Biopsy; D015816:Cell Adhesion Molecules, Neuronal / Q000235:genetics*; D045744:Cell Line, Tumor; D016000:Cluster Analysis; D003570:Cytidine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D019175:DNA Methylation; D044127:Epigenesis, Genetic / Q000187:drug effects; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D055106:Genome-Wide Association Study; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D009362:Neoplasm Metastasis; D009479:Neuropeptides / Q000235:genetics*; D011401:Promoter Regions, Genetic; D000091363:Protocadherins; D011956:Receptors, Cell Surface / Q000235:genetics*; D012208:Rhabdomyosarcoma / Q000235:genetics* / Q000473:pathology*; D059467:Transcriptome
chemical_list: D015816:Cell Adhesion Molecules, Neuronal; D006877:Hydroxamic Acids; D009479:Neuropeptides; C484820:PCDHA4 protein, human; D000091363:Protocadherins; D011956:Receptors, Cell Surface; C012589:trichostatin A; D003570:Cytidine Triphosphate; C038673:5-aza-2'-deoxycytidine-5'-triphosphate; D001374:Azacitidine
keywords: DNA methylation; Epigenetics; Microarray; PCDHA4; Rhabdomyosarcoma