Genistein blocks breast cancer cells in the G 2M phase of the cell cycle

V. Cappelletti, L. Fioravanti, P. Miodini, G. Di Fronzo

Research output: Contribution to journalArticle

Abstract

Genistein, a natural isoflavone phytoestrogen present in soybeans, caused a dose-dependent growth inhibition of the two hormone-sensitive cell lines T47D and ZR75.1 and of the two hormone-independent cell lines MDAMB-231 and BT20. Flow cytometric analysis of cells treated for 4 days with 15 and 30 μM genistein showed a dose-dependent accumulation in the G 2M phase of the cell cycle. At the highest tested concentration, there was a sevenfold increase in the percentage of cells in G 2M (63%) with respect to the control (9%) in the case of T47D cells and a 2.4-fold increase in the case of BT20. An intermediate fourfold accumulation was observed in the case of MDAMB-231 and ZR75.1. The G 2M arrest was coupled with a parallel depletion of the G 0/G 1 phase. To understand the mechanism of action underlying the block in G 2M induced by genistein, we investigated the expression and the activity of cyclins and of cyclin-dependent kinases specifically involved in the G 2→M transition. As expected, p34(cdc-2) expression, monitored by Western blotting, was unaffected by genistein treatment in all cell lines. With exception of the T47D cell line, we revealed an increase in the tyrosine phosphorylated form of p34, suggesting an inactivation of the p34(cdc-2) catalytic activity consequent to treatment of cells with genistein. In fact, immunoprecipitates from genistein-treated MDAMB-231 and BT20 cells displayed a fourfold decrease in kinase activity evaluated using the histone H1 as substrate. Conversely, no variation in kinase activity was observed between treated and untreated ZR75.1 cells despite the increase in p34 phosphorylation. In cells treated with 30 μM genistein, cyclin B 1 (p62) increased 2.8-,8-and 103-fold, respectively, in BT20, MDAMB-231, and ZR75.1 cells, suggesting an accumulation of the p62, which is instead rapidly degraded in cycling cells. No effects were observed on cyclin expression in T47D cells. We therefore conclude that genistein causes a G 2M arrest in breast cancer cell lines, but that such growth arrest is not necessarily coupled with deregulation of the p34(cdc-2)/cyclin B 1 complex only in all of the studied cell lines. (C) 2000 Wiley-Liss, Inc.

Original languageEnglish
Pages (from-to)594-600
Number of pages7
JournalJournal of Cellular Biochemistry
Volume79
Issue number4
DOIs
Publication statusPublished - 2000

Fingerprint

Gastrin-Secreting Cells
Genistein
Cell Cycle
Cells
Breast Neoplasms
Cell Line
Cyclin B
Cyclins
Phosphotransferases
Hormones
Phytoestrogens
Phosphorylation
Isoflavones
Deregulation
Cyclin-Dependent Kinases
Histones
Growth
Tyrosine
Soybeans
Catalyst activity

Keywords

  • Breast cancer
  • G M phase
  • Genistein

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

Genistein blocks breast cancer cells in the G 2M phase of the cell cycle. / Cappelletti, V.; Fioravanti, L.; Miodini, P.; Di Fronzo, G.

In: Journal of Cellular Biochemistry, Vol. 79, No. 4, 2000, p. 594-600.

Research output: Contribution to journalArticle

Cappelletti, V. ; Fioravanti, L. ; Miodini, P. ; Di Fronzo, G. / Genistein blocks breast cancer cells in the G 2M phase of the cell cycle. In: Journal of Cellular Biochemistry. 2000 ; Vol. 79, No. 4. pp. 594-600.
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PY - 2000

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N2 - Genistein, a natural isoflavone phytoestrogen present in soybeans, caused a dose-dependent growth inhibition of the two hormone-sensitive cell lines T47D and ZR75.1 and of the two hormone-independent cell lines MDAMB-231 and BT20. Flow cytometric analysis of cells treated for 4 days with 15 and 30 μM genistein showed a dose-dependent accumulation in the G 2M phase of the cell cycle. At the highest tested concentration, there was a sevenfold increase in the percentage of cells in G 2M (63%) with respect to the control (9%) in the case of T47D cells and a 2.4-fold increase in the case of BT20. An intermediate fourfold accumulation was observed in the case of MDAMB-231 and ZR75.1. The G 2M arrest was coupled with a parallel depletion of the G 0/G 1 phase. To understand the mechanism of action underlying the block in G 2M induced by genistein, we investigated the expression and the activity of cyclins and of cyclin-dependent kinases specifically involved in the G 2→M transition. As expected, p34(cdc-2) expression, monitored by Western blotting, was unaffected by genistein treatment in all cell lines. With exception of the T47D cell line, we revealed an increase in the tyrosine phosphorylated form of p34, suggesting an inactivation of the p34(cdc-2) catalytic activity consequent to treatment of cells with genistein. In fact, immunoprecipitates from genistein-treated MDAMB-231 and BT20 cells displayed a fourfold decrease in kinase activity evaluated using the histone H1 as substrate. Conversely, no variation in kinase activity was observed between treated and untreated ZR75.1 cells despite the increase in p34 phosphorylation. In cells treated with 30 μM genistein, cyclin B 1 (p62) increased 2.8-,8-and 103-fold, respectively, in BT20, MDAMB-231, and ZR75.1 cells, suggesting an accumulation of the p62, which is instead rapidly degraded in cycling cells. No effects were observed on cyclin expression in T47D cells. We therefore conclude that genistein causes a G 2M arrest in breast cancer cell lines, but that such growth arrest is not necessarily coupled with deregulation of the p34(cdc-2)/cyclin B 1 complex only in all of the studied cell lines. (C) 2000 Wiley-Liss, Inc.

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