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Global Demethylation of Rat Chondrosarcoma Cells after Treatment with 5-Aza-2′-Deoxycytidine Results in Increased Tumorigenicity

Abnormal patterns of DNA methylation are observed in several types of human cancer. While localized DNA methylation of CpG islands has been associated with gene silencing, the effect that genome-wide loss of methylation has on tumorigenesis is not completely known. To examine its effect on tumorigen...

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Autores principales: Hamm, Christopher A., Xie, Hehuang, Costa, Fabricio F., Vanin, Elio F., Seftor, Elisabeth A., Sredni, Simone T., Bischof, Jared, Wang, Deli, Bonaldo, Maria F., Hendrix, Mary J. C., Soares, Marcelo B.
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2790612/
https://www.ncbi.nlm.nih.gov/pubmed/20019818
http://dx.doi.org/10.1371/journal.pone.0008340
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author Hamm, Christopher A.
Xie, Hehuang
Costa, Fabricio F.
Vanin, Elio F.
Seftor, Elisabeth A.
Sredni, Simone T.
Bischof, Jared
Wang, Deli
Bonaldo, Maria F.
Hendrix, Mary J. C.
Soares, Marcelo B.
author_facet Hamm, Christopher A.
Xie, Hehuang
Costa, Fabricio F.
Vanin, Elio F.
Seftor, Elisabeth A.
Sredni, Simone T.
Bischof, Jared
Wang, Deli
Bonaldo, Maria F.
Hendrix, Mary J. C.
Soares, Marcelo B.
author_sort Hamm, Christopher A.
collection PubMed
description Abnormal patterns of DNA methylation are observed in several types of human cancer. While localized DNA methylation of CpG islands has been associated with gene silencing, the effect that genome-wide loss of methylation has on tumorigenesis is not completely known. To examine its effect on tumorigenesis, we induced DNA demethylation in a rat model of human chondrosarcoma using 5-aza-2-deoxycytidine. Rat specific pyrosequencing assays were utilized to assess the methylation levels in both LINEs and satellite DNA sequences following 5-aza-2-deoxycytidine treatment. Loss of DNA methylation was accompanied by an increase in invasiveness of the rat chondrosarcoma cells, in vitro, as well as by an increase in tumor growth in vivo. Subsequent microarray analysis provided insight into the gene expression changes that result from 5-aza-2-deoxycytidine induced DNA demethylation. In particular, two genes that may function in tumorigenesis, sox-2 and midkine, were expressed at low levels in control cells but upon 5-aza-2-deoxycytidine treatment these genes became overexpressed. Promoter region DNA analysis revealed that these genes were methylated in control cells but became demethylated following 5-aza-2-deoxycytidine treatment. Following withdrawal of 5-aza-2-deoxycytidine, the rat chondrosarcoma cells reestablished global DNA methylation levels that were comparable to that of control cells. Concurrently, invasiveness of the rat chondrosarcoma cells, in vitro, decreased to a level indistinguishable to that of control cells. Taken together these experiments demonstrate that global DNA hypomethylation induced by 5-aza-2-deoxycytidine may promote specific aspects of tumorigenesis in rat chondrosarcoma cells.
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spelling pubmed-27906122009-12-18 Global Demethylation of Rat Chondrosarcoma Cells after Treatment with 5-Aza-2′-Deoxycytidine Results in Increased Tumorigenicity Hamm, Christopher A. Xie, Hehuang Costa, Fabricio F. Vanin, Elio F. Seftor, Elisabeth A. Sredni, Simone T. Bischof, Jared Wang, Deli Bonaldo, Maria F. Hendrix, Mary J. C. Soares, Marcelo B. PLoS One Research Article Abnormal patterns of DNA methylation are observed in several types of human cancer. While localized DNA methylation of CpG islands has been associated with gene silencing, the effect that genome-wide loss of methylation has on tumorigenesis is not completely known. To examine its effect on tumorigenesis, we induced DNA demethylation in a rat model of human chondrosarcoma using 5-aza-2-deoxycytidine. Rat specific pyrosequencing assays were utilized to assess the methylation levels in both LINEs and satellite DNA sequences following 5-aza-2-deoxycytidine treatment. Loss of DNA methylation was accompanied by an increase in invasiveness of the rat chondrosarcoma cells, in vitro, as well as by an increase in tumor growth in vivo. Subsequent microarray analysis provided insight into the gene expression changes that result from 5-aza-2-deoxycytidine induced DNA demethylation. In particular, two genes that may function in tumorigenesis, sox-2 and midkine, were expressed at low levels in control cells but upon 5-aza-2-deoxycytidine treatment these genes became overexpressed. Promoter region DNA analysis revealed that these genes were methylated in control cells but became demethylated following 5-aza-2-deoxycytidine treatment. Following withdrawal of 5-aza-2-deoxycytidine, the rat chondrosarcoma cells reestablished global DNA methylation levels that were comparable to that of control cells. Concurrently, invasiveness of the rat chondrosarcoma cells, in vitro, decreased to a level indistinguishable to that of control cells. Taken together these experiments demonstrate that global DNA hypomethylation induced by 5-aza-2-deoxycytidine may promote specific aspects of tumorigenesis in rat chondrosarcoma cells. Public Library of Science 2009-12-17 /pmc/articles/PMC2790612/ /pubmed/20019818 http://dx.doi.org/10.1371/journal.pone.0008340 Text en Hamm et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Hamm, Christopher A.
Xie, Hehuang
Costa, Fabricio F.
Vanin, Elio F.
Seftor, Elisabeth A.
Sredni, Simone T.
Bischof, Jared
Wang, Deli
Bonaldo, Maria F.
Hendrix, Mary J. C.
Soares, Marcelo B.
Global Demethylation of Rat Chondrosarcoma Cells after Treatment with 5-Aza-2′-Deoxycytidine Results in Increased Tumorigenicity
title Global Demethylation of Rat Chondrosarcoma Cells after Treatment with 5-Aza-2′-Deoxycytidine Results in Increased Tumorigenicity
title_full Global Demethylation of Rat Chondrosarcoma Cells after Treatment with 5-Aza-2′-Deoxycytidine Results in Increased Tumorigenicity
title_fullStr Global Demethylation of Rat Chondrosarcoma Cells after Treatment with 5-Aza-2′-Deoxycytidine Results in Increased Tumorigenicity
title_full_unstemmed Global Demethylation of Rat Chondrosarcoma Cells after Treatment with 5-Aza-2′-Deoxycytidine Results in Increased Tumorigenicity
title_short Global Demethylation of Rat Chondrosarcoma Cells after Treatment with 5-Aza-2′-Deoxycytidine Results in Increased Tumorigenicity
title_sort global demethylation of rat chondrosarcoma cells after treatment with 5-aza-2′-deoxycytidine results in increased tumorigenicity
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2790612/
https://www.ncbi.nlm.nih.gov/pubmed/20019818
http://dx.doi.org/10.1371/journal.pone.0008340
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