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Seven cancer related genes (<jats:italic>THBS-1, CASP8, HIN-1, TIG-1, BLU, SPARC<\/jats:italic>, and<jats:italic>HIC-1<\/jats:italic>) that have been shown to have epigenetic changes in adult cancers and play important roles in the regulation of angiogenesis, tumor growth, and apoptosis were analyzed to investigate the role epigenetic alterations play in determining NB phenotype.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Two NB cell lines (tumorigenic LA1-55n and non-tumorigenic LA1-5s) that differ in their ability to form colonies in soft agar and tumors in nude mice were used. Quantitative RNA expression analyses were performed on seven genes in LA1-5s, LA1-55n and 5-Aza-dC treated LA1-55n NB cell lines. The methylation status around<jats:italic>THBS-1, HIN-1, TIG-1<\/jats:italic>and<jats:italic>CASP8<\/jats:italic>promoters was examined using methylation specific PCR. Chromatin immunoprecipitation assay was used to examine histone modifications along the<jats:italic>THBS-1<\/jats:italic>promoter. Luciferase assay was used to determine<jats:italic>THBS-1<\/jats:italic>promoter activity. Cell proliferation assay was used to examine the effect of 5-Aza-dC on NB cell growth. The soft agar assay was used to determine the tumorigenicity.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Promoter methylation values for<jats:italic>THBS-1<\/jats:italic>,<jats:italic>HIN-1<\/jats:italic>,<jats:italic>TIG-1<\/jats:italic>, and<jats:italic>CASP8<\/jats:italic>were higher in LA1-55n cells compared to LA1-5s cells. Consistent with the promoter methylation status, lower levels of gene expression were detected in the LA1-55n cells. Histone marks associated with repressive chromatin states (H3K9Me3, H3K27Me3, and H3K4Me3) were identified in the<jats:italic>THBS-1<\/jats:italic>promoter region in the LA1-55n cells, but not the LA1-5s cells. In contrast, the three histone codes associated with an active chromatin state (acetyl H3, acetyl H4, and H3K4Me3) were present in the<jats:italic>THBS-1<\/jats:italic>promoter region in LA1-5s cells, but not the LA1-55n cells, suggesting that an accessible chromatin structure is important for<jats:italic>THBS-1<\/jats:italic>expression. We also show that 5-Aza-dC treatment of LA1-55n cells alters the DNA methylation status and the histone code in the<jats:italic>THBS-1<\/jats:italic>promoter modifies cell morphology, and inhibits their ability to form colonies in soft agar.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Our results suggest that epigenetic aberrations contribute to NB phenotype, and that tumorigenic properties can be inhibited by reversing the epigenetic changes with 5-Aza-dC.<\/jats:p><\/jats:sec>","DOI":"10.1186\/1471-2407-10-286","type":"journal-article","created":{"date-parts":[[2010,6,15]],"date-time":"2010-06-15T06:15:54Z","timestamp":1276582554000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":24,"title":["Epigenetic alterations differ in phenotypically distinct human neuroblastoma cell lines"],"prefix":"10.1186","volume":"10","author":[{"given":"Qiwei","family":"Yang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yufeng","family":"Tian","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kelly R","family":"Ostler","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandre","family":"Chlenski","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa J","family":"Guerrero","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helen R","family":"Salwen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lucy A","family":"Godley","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susan L","family":"Cohn","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2010,6,14]]},"reference":[{"key":"2085_CR1","doi-asserted-by":"publisher","first-page":"2106","DOI":"10.1016\/S0140-6736(07)60983-0","volume":"369","author":"JM Maris","year":"2007","unstructured":"Maris JM, Hogarty MD, Bagatell R, Cohn SL: Neuroblastoma. 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