Cell cycle block and apoptosis induction in a human melanoma cell line following treatment with 2-methoxyoestradiol

Therapeutic implications?

Rita Ghosh, Ann M. Ott, Divya Seetharam, Thomas J Slaga, Addanki P Kumar

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Due to minimal success with non-surgical treatment options for melanoma, it is imperative that other compounds be tested for potential preventive/therapeutic use. We have tested the ability of the endogenous oestrogenic metabolite 2-methoxyestradiol (2-ME) to inhibit the growth of human melanoma cells in culture. 2-ME inhibited the growth of all the melanoma cells tested, without inhibiting the growth of non-tumorigenic cells. Microscopic observations showed that treated cells exhibit the characteristic features of apoptosis. Examination of the molecular mechanism in WM98-1 cells, using biochemical assays such as a modified TUNEL staining and DNA fragmentation, confirmed the induction of apoptosis following 2-ME treatment. Flow cytometry analysis showed that, following treatment, cells are arrested in the G2/M phase of the cell cycle. Western blot analysis of the G2/M regulatory proteins suggests that cdc2 is involved in the cell cycle block by Myt1 phosphorylation following 2-ME treatment. Furthermore, examination of the levels of apoptosis regulatory proteins showed that, while levels of p53, Bax and p21 are higher, that of anti-apoptotic Bcl-2 is undetectable in cells treated with 2-ME compared with untreated controls. Taken together these results have major implications for the use of 2-ME for melanoma management. Melanoma Res 13:119-127

Original languageEnglish (US)
Pages (from-to)119-127
Number of pages9
JournalMelanoma Research
Volume13
Issue number2
DOIs
StatePublished - Apr 2003
Externally publishedYes

Fingerprint

Melanoma
Cell Cycle
Apoptosis
Cell Line
Therapeutics
Growth
CDC2 Protein Kinase
Apoptosis Regulatory Proteins
G2 Phase
In Situ Nick-End Labeling
Therapeutic Uses
DNA Fragmentation
Cell Division
2-methoxyestradiol
Flow Cytometry
Cell Culture Techniques
Western Blotting
Phosphorylation
Staining and Labeling

Keywords

  • 2-methoxyoestradiol
  • Apoptosis
  • Cell cycle
  • Melanoma
  • Proliferation

ASJC Scopus subject areas

  • Cancer Research
  • Dermatology

Cite this

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title = "Cell cycle block and apoptosis induction in a human melanoma cell line following treatment with 2-methoxyoestradiol: Therapeutic implications?",
abstract = "Due to minimal success with non-surgical treatment options for melanoma, it is imperative that other compounds be tested for potential preventive/therapeutic use. We have tested the ability of the endogenous oestrogenic metabolite 2-methoxyestradiol (2-ME) to inhibit the growth of human melanoma cells in culture. 2-ME inhibited the growth of all the melanoma cells tested, without inhibiting the growth of non-tumorigenic cells. Microscopic observations showed that treated cells exhibit the characteristic features of apoptosis. Examination of the molecular mechanism in WM98-1 cells, using biochemical assays such as a modified TUNEL staining and DNA fragmentation, confirmed the induction of apoptosis following 2-ME treatment. Flow cytometry analysis showed that, following treatment, cells are arrested in the G2/M phase of the cell cycle. Western blot analysis of the G2/M regulatory proteins suggests that cdc2 is involved in the cell cycle block by Myt1 phosphorylation following 2-ME treatment. Furthermore, examination of the levels of apoptosis regulatory proteins showed that, while levels of p53, Bax and p21 are higher, that of anti-apoptotic Bcl-2 is undetectable in cells treated with 2-ME compared with untreated controls. Taken together these results have major implications for the use of 2-ME for melanoma management. Melanoma Res 13:119-127",
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AU - Kumar, Addanki P

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