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  • Title: Effect of 5-azacytidine on DNA methylation and the malignant properties of B16 melanoma cells.
    Author: Trainer DL, Kline T, Mallon F, Greig R, Poste G.
    Journal: Cancer Res; 1985 Dec; 45(12 Pt 1):6124-30. PubMed ID: 2415238.
    Abstract:
    The role of DNA methylation in the expression of the metastatic phenotype in B16 murine melanoma cells in syngeneic C57BL/6 mice has been investigated. B16 cultures were incubated in vitro for either 6 or 18 h with the DNA hypomethylating agents, 5-azacytidine (5-Aza-CR) or 5-fluoro-2'-deoxycytidine (FCdR). At various times (1-13 days) following treatment, tumor cells were tested for their ability to form metastatic deposits when injected at different doses either i.v. (experimental metastasis) or s.c. in the footpad (spontaneous metastasis). Both 5-Aza-CR (0.5-15 microM) and FCdR (0.3-30 microM) caused a dose-dependent increase in the ability of B16 cells to form experimental pulmonary metastases. Increased capacity to form experimental pulmonary metastases was evident 24 h following treatment with 5-Aza-CR and 13 days following treatment with FCdR. The enhanced metastatic burden involved both an increase in the median number of lung colonies and a substantial increase in the size of individual lesions. 5-Aza-CR or FCdR treatment of B16 cell populations did not influence either the tumorigenicity or their ability to form spontaneous metastases. Parallel in vitro experiments using high-performance liquid chromatography analysis of cellular DNA demonstrated that under conditions in which 5-Aza-CR and FCdR enhanced formation of experimental metastases by B16 cells, there were readily detectable alterations in the 5-methylcytosine levels in DNA extracted from drug-treated cultures. These data suggest that drug-induced alterations in DNA methylation can affect biochemical pathway(s) whose expression is associated with the successful organ colonization by circulating tumor cells.
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