Paternal combined botanicals contribute to the prevention of estrogen receptor-negative mammary cancer in transgenic mice.BACKGROUND
 Parental nutritional interventions have considerably affected gametogenesis and embryogenesis, leading to differential susceptibility of offspring to chronic diseases like cancer.  Moreover, combinatorial bioactive diets are more efficacious in ameliorating epigenetic aberrations in tumorigenesis.


 OBJECTIVE
 We sought to investigate the transgenerational influence and epigenetic regulation of paternal sulforaphane (SFN)-rich broccoli sprouts (BSp) and epigallocatechin-3-gallate (EGCG)-rich green tea polyphenols (GTPs) consumption on the prevention of estrogen receptor-negative [ER(-)] mammary cancer with transgenic mice.


 Human breast cancer (BC) cells were used to detect cell viability and epigenetic-related gene expression after treatment with EGCG or/and SFN.  Twenty-four C3 or HER2/neu males were randomly assigned into four groups and treated with: control, 26% BSp (w/w) in food, 0.5% GTPs (w/v) in drinking water or combined BSp and GTPs for 7 wk before mating.  Tumor growth of non-treated female pups was monitored weekly for 19 wk (C3) and 25 wk (HER2/neu).  Tumor- and epigenetic-related protein expression and enzyme activities in mammary tumors were measured.  EGCG and SFN inhibited BC cell growth via epigenetic regulations.  Combined BSp and GTPs synergistically (CIn < 1) suppressed tumor growth over time (P < 0.001) in two mouse models.  Key tumor-related proteins were found differentially expressed (P < 0.05) along with epigenetic regulations in offspring mammary tumors.  The transcriptome profile of sperms derived from dietary-treated males revealed differentially expressed genes correlated with spermatogenesis and breast cancer progression.  DNA methylomes of the sperm and further integrated analysis with transcriptome indicate that DNA methylation alone may not contribute to sufficient regulation in dietary-treated sperm pronucleus leading to offspring tumor suppression.


 CONCLUSION
 Collectively, paternal consumption of combined BSp and GTPs shows potential for preventing ER(-) mammary cancer through transgenerational effects.
C