Mechanism for the suppression of the mammalian stress response by genistein, an anticancer phytoestrogen from soy.
BACKGROUND
Soy products contain high levels of genistein, a phytoestrogen that is a potent inhibitor of cell proliferation and angiogenesis. Genistein has been found to inhibit the growth of carcinogen-induced cancers in rats and human leukemia cells transplanted into mice. The induction of stress proteins (e.g., glucose-related proteins and heat shock proteins) in tumor cells has been shown to protect them against programmed cell death; this stress response is inhibited by genistein. The mechanism(s) by which genistein affects certain stress response genes was explored in this study.


METHODS
Mammalian cell cultures were treated with azetidine, a proline analog, which elicits a stress response that includes the induction of the expression of glucose-regulated protein GRP78 and heat shock protein HSP70. The effects of azetidine and/or genistein treatment on cellular levels of grp78 and hsp70 messenger RNAs and proteins were measured by northern blot hybridization and western blot analyses, respectively, and the binding of nuclear factors to sequence motifs in the upstream (promoter) regions of these two genes were examined by electrophoretic mobility shift assays.


RESULTS
Genistein antagonized the binding of a specific transcription factor, nuclear factor-Y/CCAAT binding factor (NF-Y/CBF), to the CCAAT sequence element most proximal to the transcription start sites in the hsp70 and grp78 promoters; this CCAAT element was previously shown to be necessary for full-stress inducibility of both genes. Treatment of cells with genistein converted NF-Y/CBF into a nonbinding, transcriptionally inactive form.


CONCLUSIONS
The anticancer effects of genistein may be related to its ability to reduce the expression of stress response-related genes.
Zhou
Y
Y
Department of Biochemistry and Molecular Biology, USC/Norris Comprehensive Cancer Center, University of Southern California School of Medicine, Los Angeles 90033, USA.
Lee
A S
AS
D000818:Animals; D016588:Anticarcinogenic Agents / Q000494:pharmacology*; D001384:Azetidines / Q000494:pharmacology; D015152:Blotting, Northern; D015153:Blotting, Western; D002352:Carrier Proteins / Q000032:analysis* / Q000235:genetics; D006224:Cricetinae; D003412:Cricetulus; D000091342:Endoplasmic Reticulum Chaperone BiP; D005347:Fibroblasts; D005786:Gene Expression Regulation / Q000187:drug effects; D019833:Genistein / Q000494:pharmacology*; D018840:HSP70 Heat-Shock Proteins / Q000032:analysis* / Q000235:genetics; D006360:Heat-Shock Proteins; D008168:Lung / Q000166:cytology; D051379:Mice; D018832:Molecular Chaperones / Q000032:analysis* / Q000235:genetics; D009154:Mutation
D016588:Anticarcinogenic Agents; D001384:Azetidines; D002352:Carrier Proteins; D000091342:Endoplasmic Reticulum Chaperone BiP; D018840:HSP70 Heat-Shock Proteins; C000717247:HSPA5 protein, human; D006360:Heat-Shock Proteins; C000717248:Hspa5 protein, mouse; D018832:Molecular Chaperones; C082735:azetidine; D019833:Genistein