Effect of 5-azacytidine on DNA methylation and the malignant properties of B16 melanoma cells

D. L. Trainer, T. Kline, F. Mallon, R. Greig, George Poste

Research output: Contribution to journalArticle

26 Citations (Scopus)

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 μM) and FCdR (0.3-30 μM) 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.

Original languageEnglish (US)
Pages (from-to)6124-6130
Number of pages7
JournalCancer Research
Volume45
Issue number12
StatePublished - 1985
Externally publishedYes

Fingerprint

Azacitidine
Experimental Melanomas
DNA Methylation
Neoplasm Metastasis
Lung
DNA
5-Methylcytosine
Circulating Neoplastic Cells
Inbred C57BL Mouse
Pharmaceutical Preparations
High Pressure Liquid Chromatography
Phenotype
Population
Neoplasms

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Effect of 5-azacytidine on DNA methylation and the malignant properties of B16 melanoma cells. / Trainer, D. L.; Kline, T.; Mallon, F.; Greig, R.; Poste, George.

In: Cancer Research, Vol. 45, No. 12, 1985, p. 6124-6130.

Research output: Contribution to journalArticle

Trainer, DL, Kline, T, Mallon, F, Greig, R & Poste, G 1985, 'Effect of 5-azacytidine on DNA methylation and the malignant properties of B16 melanoma cells', Cancer Research, vol. 45, no. 12, pp. 6124-6130.
Trainer, D. L. ; Kline, T. ; Mallon, F. ; Greig, R. ; Poste, George. / Effect of 5-azacytidine on DNA methylation and the malignant properties of B16 melanoma cells. In: Cancer Research. 1985 ; Vol. 45, No. 12. pp. 6124-6130.
@article{566a894fe2574cf4ae43d36803d1a019,
title = "Effect of 5-azacytidine on DNA methylation and the malignant properties of B16 melanoma cells",
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 μM) and FCdR (0.3-30 μM) 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.",
author = "Trainer, {D. L.} and T. Kline and F. Mallon and R. Greig and George Poste",
year = "1985",
language = "English (US)",
volume = "45",
pages = "6124--6130",
journal = "Cancer Research",
issn = "0008-5472",
publisher = "American Association for Cancer Research Inc.",
number = "12",

}

TY - JOUR

T1 - Effect of 5-azacytidine on DNA methylation and the malignant properties of B16 melanoma cells

AU - Trainer, D. L.

AU - Kline, T.

AU - Mallon, F.

AU - Greig, R.

AU - Poste, George

PY - 1985

Y1 - 1985

N2 - 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 μM) and FCdR (0.3-30 μM) 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.

AB - 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 μM) and FCdR (0.3-30 μM) 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.

UR - http://www.scopus.com/inward/record.url?scp=0022359321&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0022359321&partnerID=8YFLogxK

M3 - Article

VL - 45

SP - 6124

EP - 6130

JO - Cancer Research

JF - Cancer Research

SN - 0008-5472

IS - 12

ER -