TY - JOUR
T1 - Epstein-Barr virus-specific methylation of human genes in gastric cancer cells
AU - Ryan, Julie L.
AU - Jones, Richard J.
AU - Kenney, Shannon C.
AU - Rivenbark, Ashley G.
AU - Tang, Weihua
AU - Knight, Elizabeth Rw
AU - Coleman, William B.
AU - Gulley, Margaret L.
N1 - Funding Information:
The authors thank Hongxin Fan, MD for developing the BamH1W EBV viral load assay, and Dr. Henri J. Delecluse for providing the recombinant EBV strain. This study was sponsored by grants from the National Cancer Institute (R01CA058853 and T32-ES07017) and by the University of North Carolina Department of Pathology and Laboratory Medicine. We are also grateful for support from the University Cancer Research Fund of the Lineberger Comprehensive Cancer Center, the NC Translational and Clinical Sciences Institute, and the Translational Pathology Laboratory, Genomics and Bioinformatics Core Facility, Nucleic Acids Core Facility, Genome Analysis Facility, and the GI SPORE group (P50 CA106991) of the University of North Carolina at Chapel Hill.
PY - 2010
Y1 - 2010
N2 - Background. Epstein-Barr Virus (EBV) is found in 10% of all gastric adenocarcinomas but its role in tumor development and maintenance remains unclear. The objective of this study was to examine EBV-mediated dysregulation of cellular factors implicated in gastric carcinogenesis. Methods. Gene expression patterns were examined in EBV-negative and EBV-positive AGS gastric epithelial cells using a low density microarray, reverse transcription PCR, histochemical stains, and methylation-specific DNA sequencing. Expression of PTGS2 (COX2) was measured in AGS cells and in primary gastric adenocarcinoma tissues. Results. In array studies, nearly half of the 96 human genes tested, representing 15 different cancer-related signal transduction pathways, were dysregulated after EBV infection. Reverse transcription PCR confirmed significant impact on factors having diverse functions such as cell cycle regulation (IGFBP3, CDKN2A, CCND1, HSP70, ID2, ID4), DNA repair (BRCA1, TFF1), cell adhesion (ICAM1), inflammation (COX2), and angiogenesis (HIF1A). Demethylation using 5-aza-2'-deoxycytidine reversed the EBV-mediated dysregulation for all 11 genes listed here. For some promoter sequences, CpG island methylation and demethylation occurred in an EBV-specific pattern as shown by bisulfite DNA sequencing. Immunohistochemistry was less sensitive than was western blot for detecting downregulation of COX2 upon EBV infection. Virus-related dysregulation of COX2 levels in vitro was not recapitulated in vivo among naturally infected gastric cancer tissues. Conclusions. EBV alters human gene expression in ways that could contribute to the unique pathobiology of virus-associated cancer. Furthermore, the frequency and reversability of methylation-related transcriptional alterations suggest that demethylating agents have therapeutic potential for managing EBV-related carcinoma.
AB - Background. Epstein-Barr Virus (EBV) is found in 10% of all gastric adenocarcinomas but its role in tumor development and maintenance remains unclear. The objective of this study was to examine EBV-mediated dysregulation of cellular factors implicated in gastric carcinogenesis. Methods. Gene expression patterns were examined in EBV-negative and EBV-positive AGS gastric epithelial cells using a low density microarray, reverse transcription PCR, histochemical stains, and methylation-specific DNA sequencing. Expression of PTGS2 (COX2) was measured in AGS cells and in primary gastric adenocarcinoma tissues. Results. In array studies, nearly half of the 96 human genes tested, representing 15 different cancer-related signal transduction pathways, were dysregulated after EBV infection. Reverse transcription PCR confirmed significant impact on factors having diverse functions such as cell cycle regulation (IGFBP3, CDKN2A, CCND1, HSP70, ID2, ID4), DNA repair (BRCA1, TFF1), cell adhesion (ICAM1), inflammation (COX2), and angiogenesis (HIF1A). Demethylation using 5-aza-2'-deoxycytidine reversed the EBV-mediated dysregulation for all 11 genes listed here. For some promoter sequences, CpG island methylation and demethylation occurred in an EBV-specific pattern as shown by bisulfite DNA sequencing. Immunohistochemistry was less sensitive than was western blot for detecting downregulation of COX2 upon EBV infection. Virus-related dysregulation of COX2 levels in vitro was not recapitulated in vivo among naturally infected gastric cancer tissues. Conclusions. EBV alters human gene expression in ways that could contribute to the unique pathobiology of virus-associated cancer. Furthermore, the frequency and reversability of methylation-related transcriptional alterations suggest that demethylating agents have therapeutic potential for managing EBV-related carcinoma.
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U2 - 10.1186/1750-9378-5-27
DO - 10.1186/1750-9378-5-27
M3 - Article
C2 - 21194482
AN - SCOPUS:78650644411
SN - 1750-9378
VL - 5
JO - Infectious Agents and Cancer
JF - Infectious Agents and Cancer
IS - 1
M1 - 27
ER -