Methylation analysis of EDNRB in human colon tissues of Hirschsprung's disease

Pediatr Surg Int. 2013 Jul;29(7):683-8. doi: 10.1007/s00383-013-3308-6. Epub 2013 Apr 12.

Abstract

Purpose: Hirschsprung's disease (HSCR) is characterized by absence of the enteric nervous system in a variable portion of the distal gut. The endothelin receptor type B (EDNRB) gene has been localized to the chromosome 13q22 region and encodes a G-protein coupled receptor, is generally accepted as a crucial gene for HSCR. This study is to identify the epigenetic changes of EDNRB in the pathogenesis of HSCR.

Methods: We investigated the expression levels of EDNRB in 58 HSCR patients and 25 unrelated controls, using reverse transcriptase polymerase chain reaction (RT-PCR) and western blot assay. Moreover, using the methylation-specific polymerase chain reaction, we examined the methylation status of the promoter region of EDNRB.

Results: Aberrant high expression level of EDNRB was detected in HSCR patients compared with the control group (P = 0.023). Besides, western blot assay confirmed the up-regulation of EDNRB in the post transcription level in the aganglionosis segment of HSCR patients. Furthermore, there was a significantly lower ratio of methylation level of EDNRB in HSCR.

Conclusions: Our study demonstrates that epigenetic inactivation of the EDNRB gene may play a role in the development of HSCR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western / methods
  • China
  • Colon / metabolism*
  • Female
  • Hirschsprung Disease / genetics*
  • Hirschsprung Disease / metabolism*
  • Humans
  • Infant
  • Male
  • Methylation
  • Polymerase Chain Reaction / methods
  • Receptor, Endothelin B / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction / methods

Substances

  • Receptor, Endothelin B