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

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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.

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Acknowledgments

We thank Dr. Xiaoqun Xu, Weiwei Jiang, Xiaofeng Lv and Changgui Lu (Nanjing Children’s Hospital Affiliated to Nanjing Medical University) for sample collection. This study was supported by Grants SKLRM-KF-1104, Nanjing Science and Technique Development Foundation (201108010) and Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

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Authors and Affiliations

  1. State Key Laboratory of Reproductive Medicine, Institute of Toxicology, School of Public Health, Nanjing Medical University, Nanjing, 210029, China

    Weibing Tang, Bo Li, Junwei Tang, Kang Liu, Jingjing Qin, Wei Wu, Xiaoqun Xu & Yankai Xia

  2. Key Laboratory of Modern Toxicology, Nanjing Medical University, Ministry of Education, Nanjing, China

    Wei Wu & Yankai Xia

  3. Department of Pediatric Surgery, Nanjing Children’s Hospital Affiliated Nanjing Medical University, Nanjing, 210008, China

    Weibing Tang, Bo Li, Junwei Tang, Kang Liu, Jingjing Qin, Qiming Geng, Jie Zhang, Huan Chen & Xiaoqun Xu

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  1. Weibing Tang
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  2. Bo Li
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  3. Junwei Tang
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Corresponding author

Correspondence to Yankai Xia.

Additional information

W. Tang, B. Li, and J. Tang contributed equally to this work.

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Tang, W., Li, B., Tang, J. et al. Methylation analysis of EDNRB in human colon tissues of Hirschsprung’s disease. Pediatr Surg Int 29, 683–688 (2013). https://doi.org/10.1007/s00383-013-3308-6

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  • DOI: https://doi.org/10.1007/s00383-013-3308-6

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