p300 expression repression by hypermethylation associated with tumour invasion and metastasis in oesophageal squamous cell carcinoma

J Clin Pathol. 2007 Nov;60(11):1249-53. doi: 10.1136/jcp.2006.044099.

Abstract

Background: Aberrant promoter methylation is an important mechanism for gene silencing.

Aims: To evaluate the promoter methylation status of p300 gene in patients with oesophageal squamous cell carcinoma (OSCC).

Methods: The methylation status of p300 promoter was analysed by methylation-specific PCR (MSP) in 50 OSCC tissues and the matching non-cancerous tissues. Oesophageal cancer cell lines (ECa-109 and TE-10) were treated with the demethylation agent 5-aza-2'-deoxycytidine (5-Aza-CdR), and p300 mRNA expression was detected by RT-PCR.

Results: p300 methylation was found in 42% (21/50) of the OSCC tissues, but in only 20% (10/50) of the corresponding non-cancerous tissues (p = 0.017). In OSCC samples, 65% of those with deep tumour invasion (adventitia) and 63% samples with metastasis revealed p300 promoter methylation (p<0.05). p300 mRNA expression was observed in 19.0% (4/21) of methylated tumours and 58.6% (17/29) of unmethylated tumours (p = 0.005). In addition, p300 mRNA expression was observed in 40% (4/10) of methylated non-neoplastic tissues and 87.5% (35/40) of unmethylated non-tumours (p = 0.001). The demethylation caused by 5-Aza-CdR increased the p300 mRNA expression levels in oesophageal cancer cell lines.

Conclusions: p300 transcription silenced by promoter hypermethylation could play a role in the pathogenesis of oesophageal squamous cell carcinoma.

MeSH terms

  • Azacitidine / analogs & derivatives
  • Azacitidine / pharmacology
  • Carcinoma, Squamous Cell / genetics
  • Carcinoma, Squamous Cell / metabolism*
  • Carcinoma, Squamous Cell / pathology
  • Carcinoma, Squamous Cell / secondary
  • DNA Methylation
  • DNA, Neoplasm / genetics
  • Decitabine
  • E1A-Associated p300 Protein / genetics
  • E1A-Associated p300 Protein / metabolism*
  • Esophageal Neoplasms / genetics
  • Esophageal Neoplasms / metabolism*
  • Esophageal Neoplasms / pathology
  • Female
  • Gene Expression Regulation, Neoplastic / drug effects
  • Gene Silencing
  • Humans
  • Lymphatic Metastasis
  • Male
  • Neoplasm Invasiveness
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism*
  • Promoter Regions, Genetic / genetics
  • RNA, Messenger / genetics
  • RNA, Neoplasm / genetics
  • Reverse Transcriptase Polymerase Chain Reaction / methods

Substances

  • DNA, Neoplasm
  • Neoplasm Proteins
  • RNA, Messenger
  • RNA, Neoplasm
  • Decitabine
  • E1A-Associated p300 Protein
  • EP300 protein, human
  • Azacitidine