title: Association between human papillomavirus and Epstein - Barr virus DNA and gene promoter methylation of RB1 and CDH1 in the cervical lesions: a transversal study.
abstract: BACKGROUND
  Human papillomavirus (HPV) inactivates the retinoblastoma 1 (RB1) gene by promoter methylation and reduces cellular E-cadherin expression by overexpression of DNA methyltransferase 1 (DNMT1). The Epstein-Barr virus (EBV) is an oncogenic virus that may be related to cervical carcinogenesis. In gastric cancer, it has been demonstrated that E-cadherin gene (CDH1) hypermethylation is associated with DNMT1 overexpression by EBV infection. Our aim was to analyze the gene promoter methylation frequency of RB1 and CDH1 and verify the association between that methylation frequency and HPV and EBV infection in cervical lesions.
  
  
  METHODS
  Sixty-five samples were obtained from cervical specimens: 15 normal cervices, 17 low-grade squamous intraepithelial lesions (LSIL), 15 high-grade squamous intraepithelial lesions (HSIL), and 18 cervical cancers. HPV and EBV DNA testing was performed by PCR, and the methylation status was verified by MSP.
  
  
  RESULTS
  HPV frequency was associated with cervical cancer cases (p = 0.005) but not EBV frequency (p = 0.732). Viral co-infection showed a statistically significant correlation with cancer (p = 0.027). No viral infection was detected in 33.3% (5/15) of controls. RB1 methylated status was associated with cancer (p = 0.009) and HPV infection (p = 0.042). CDH1 methylation was not associated with cancer (p = 0.181). Controls and LSIL samples did not show simultaneous methylation, while both genes were methylated in 27.8% (5/18) of cancer samples. In the presence of EBV, CDH1 methylation was present in 27.8% (5/18) of cancer samples. Only cancer cases presented RB1 promoter methylation in the presence of HPV and EBV (33.3%).
  
  
  CONCLUSIONS
  The methylation status of both genes increased with disease progression. With EBV, RB1 methylation was a tumor-associated event because only the cancer group presented methylated RB1 with HPV infection. HPV infection was shown to be significantly correlated with cancer conditions. The global methylation frequency was higher when HPV was present, showing its epigenetic role in cervical carcinogenesis. Nevertheless, EBV seems to be a cofactor and needs to be further investigated.
  The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1159157579149317 .
authors.affiliation: Laboratory of Molecular Pathology, Pathological Anatomy Service and Pathology Department, Clementino Fraga Filho University Hospital, Federal University of Rio de Janeiro - UFRJ, Rio de Janeiro, Brazil. maccormick.thais@gmail.com.
authors.forename: Thaís M
authors.initials: TM
authors.lastname: McCormick
authors.affiliation: Laboratory of Neuropathology, Pathological Anatomy Service and Pathology Department, Clementino Fraga Filho University Hospital, Federal University of Rio de Janeiro - UFRJ, Rio de Janeiro, Brazil. nathaliecanedo@gmail.com.
authors.forename: Nathalie H S
authors.initials: NH
authors.lastname: Canedo
authors.affiliation: Gynecology Institute, Federal University of Rio de Janeiro - UFRJ, Rio de Janeiro, Brazil. yarafurtadorj@terra.com.br.
authors.forename: Yara L
authors.initials: YL
authors.lastname: Furtado
authors.affiliation: Gynecology Institute, Federal University of Rio de Janeiro - UFRJ, Rio de Janeiro, Brazil. filomenasilveira@uol.com.br.
authors.forename: Filomena A
authors.initials: FA
authors.lastname: Silveira
authors.affiliation: Gynecology Institute, Federal University of Rio de Janeiro - UFRJ, Rio de Janeiro, Brazil. limarober@gmail.com.
authors.forename: Roberto J
authors.initials: RJ
authors.lastname: de Lima
authors.affiliation: Gynecology Institute, Federal University of Rio de Janeiro - UFRJ, Rio de Janeiro, Brazil. andrea.rosman@gmail.com.
authors.forename: Andréa D F
authors.initials: AD
authors.lastname: Rosman
authors.affiliation: Gynecology Institute, Federal University of Rio de Janeiro - UFRJ, Rio de Janeiro, Brazil. gutemberg@gineco.ufrj.br.
authors.forename: Gutemberg L
authors.initials: GL
authors.lastname: Almeida Filho
authors.affiliation: Laboratory of Molecular Pathology, Pathological Anatomy Service and Pathology Department, Clementino Fraga Filho University Hospital, Federal University of Rio de Janeiro - UFRJ, Rio de Janeiro, Brazil. gloria@gcarvalho.org.|Serviço de Anatomia Patológica, Subsolo - sala 09 (Citopatologia), Hospital Universitário Clementino Fraga Filho, UFRJ, Ilha do Fundão, Rio de Janeiro, RJ, CEP 21941-913, Brazil. gloria@gcarvalho.org.
authors.forename: Maria da Glória da C
authors.initials: Mda G
authors.lastname: Carvalho
mesh_terms: D015703:Antigens, CD; D015820:Cadherins / Q000235:genetics* / Q000378:metabolism; D016022:Case-Control Studies; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation; D004279:DNA, Viral / Q000235:genetics*; D018450:Disease Progression; D044127:Epigenesis, Genetic; D020031:Epstein-Barr Virus Infections / Q000235:genetics* / Q000821:virology; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D004854:Herpesvirus 4, Human / Q000235:genetics* / Q000472:pathogenicity; D054884:Host-Pathogen Interactions; D061809:Human Papillomavirus DNA Tests; D006801:Humans; D060787:Neoplasm Grading; D027383:Papillomaviridae / Q000235:genetics* / Q000472:pathogenicity; D030361:Papillomavirus Infections / Q000235:genetics* / Q000821:virology; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D016160:Retinoblastoma Protein / Q000235:genetics* / Q000378:metabolism; D065310:Squamous Intraepithelial Lesions of the Cervix / Q000201:enzymology / Q000235:genetics* / Q000473:pathology / Q000821:virology; D002583:Uterine Cervical Neoplasms / Q000201:enzymology / Q000235:genetics* / Q000473:pathology / Q000821:virology; D002578:Uterine Cervical Dysplasia / Q000201:enzymology / Q000235:genetics* / Q000473:pathology / Q000821:virology
chemical_list: D015703:Antigens, CD; C513384:CDH1 protein, human; D015820:Cadherins; D004279:DNA, Viral; D016160:Retinoblastoma Protein; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases; C000619380:DNMT1 protein, human