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BioMed Research International
Volume 2014 (2014), Article ID 327538, 16 pages
http://dx.doi.org/10.1155/2014/327538
Research Article

Osteoponin Promoter Controlled by DNA Methylation: Aberrant Methylation in Cloned Porcine Genome

1Department of Life Sciences, National Chung Hsing University, Taichung 402, Taiwan
2School of Medicine, College of Medicine, China Medical University, Taichung 40402, Taiwan
3Department of Otolaryngology Head and Neck Surgery, China Medical University, Taichung 40402, Taiwan
4Department of Bioresources, Da-Yeh University, Changhwa 515, Taiwan
5Department of Chinese Pharmaceutical Sciences and Chinese Medicine Resources, College of Pharmacy, China Medical University, Taichung 40402, Taiwan
6Department of Animal Science and Technology, National Taiwan University, Taipei 106, Taiwan
7Department of Animal Science and Biotechnology, Tung Hai University, Taichung 407, Taiwan
8Department of Biomedical Informatics, Asia University, Taichung 41354, Taiwan
9Department of Medical Research, Human Genetic Center, China Medical University Hospital, Taichung 40447, Taiwan
10Research Center for Chinese Medicine & Acupuncture, China Medical University, Taichung 40402, Taiwan
11Rong Hsing Research Center for Translational Medicine and the iEGG Center, National Chung Hsing University, Taichung 402, Taiwan

Received 24 February 2014; Accepted 5 March 2014; Published 2 July 2014

Academic Editor: Chung Y. Hsu

Copyright © 2014 Chih-Jie Shen et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Cloned animals usually exhibited many defects in physical characteristics or aberrant epigenetic reprogramming, especially in some important organ development. Osteoponin (OPN) is an extracellular-matrix protein involved in heart and bone development and diseases. In this study, we investigated the correlation between OPN mRNA and its promoter methylation changes by the 5-aza-dc treatment in fibroblast cell and promoter assay. Aberrant methylation of porcine OPN was frequently found in different tissues of somatic nuclear transferred cloning pigs, and bisulfite sequence data suggested that the OPN promoter region −2615 to −2239 nucleotides (nt) may be a crucial regulation DNA element. In pig ear fibroblast cell culture study, the demethylation of OPN promoter was found in dose-dependent response of 5-aza-dc treatment and followed the OPN mRNA reexpression. In cloned pig study, discrepant expression pattern was identified in several cloned pig tissues, especially in brain, heart, and ear. Promoter assay data revealed that four methylated CpG sites presenting in the −2615 to −2239 nt region cause significant downregulation of OPN promoter activity. These data suggested that methylation in the OPN promoter plays a crucial role in the regulation of OPN expression that we found in cloned pigs genome.