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Abstract
Genistein, a natural isoflavonoid phyto-oestrogen, inhibits the tyrosine kinase activity
of growth factor receptors and oncogene products, as well as the in vitro growth of some tumour cell lines. The low incidence of breast cancer in countries
with a flavonoid-rich soy-based diet and the protection afforded by soy-derived products
against experimental mammary tumours in rats suggest that genistein and other isoflavonoid
compounds may exert an anti-tumour activity. We analysed the effects of genistein
on cell number and cell cycle progression (flow cytometric analysis of propidium iodide-stained
nuclei) of human breast cancer cells (MCF-7) in vitro. Genistein produced a significant, dose-dependent inhibition of MCF-7 cell growth
with an id50 of ∼ 40 μM after 72 h of incubation. Cell cycle analysis showed a reversible G2/M arrest in cell cycle progression at 10 μM genistein concentrations, whilst a marked
fall in S-phase cell percentage associated with a persistent arrest in G2/M phase was observed in cultures treated with genistein doses equal to or greater
than 50 μM. These effects were significant at 24 h of incubation; flow cytometric
analysis at later times (48 and 72 h) revealed a population of cells with decreased
DNA content and nuclear fragmentation characteristic of apoptosis. Thus, the growth
inhibitory activity of innistein in MCF-7 cells results from the sum of cytostatic
and apoptotic effects. Since the mitogenic action of insulin and insulin-like growth
factor (IGF)-I in MCF-7 cells is a tyrosine kinase-dependent phenomenon, we analysed
the genistein impact on S-phase entry produced by insulin in cultures partially synchronised
in G0/G1 phase by serum deprivation. Insulin addition after a 36-h culture period in serum-free
medium produced a strong increase in the percentage of S-phase cells (from 18.4 ±
2.3 to 46.2 ± 4.1 after 24 h) which was almost completely blocked by 100 μM genistein
(20.1 ± 3.1). Immunofluorescence analysis with a fluoresceine isothiocyanate (FITC)-conjugated
anti-phosphotyrosine antibody revealed a strong increase in MCF-7 cell staining after
insulin stimulation, but not when genistein was added with insulin. In conclusion,
the dietary phyto-oestrogen genistein inhibits in vitro growth of MCF-7 human breast cancer cells through blocks in the “critical checkpoints”
of cell cycle control and induction of apoptosis. These effects are likely to depend
on impairment in the signal transduction pathway from tyrosine kinase receptor(s).
Keywords
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Article info
Publication history
Accepted:
May 6,
1994
Received:
April 25,
1994
Identification
Copyright
© 1994 Published by Elsevier Inc.
