Increased consumption of cruciferous vegetables is associated with a reduced risk of developing prostate cancer. Indole-3-carbinol (I3C) and 3,3′-diindolylmethane (DIM) are phytochemicals derived from cruciferous vegetables that have shown promise in inhibiting prostate cancer in experimental models. Histone deacetylase (HDAC) inhibition is an emerging target for cancer prevention and therapy. We sought to examine the effects of I3C and DIM on HDACs in human prostate cancer cell lines: androgen insensitive PC-3cells and androgen sensitive LNCaP cells. I3C modestly inhibited HDAC activity in LNCaP cells by 25% but no inhibition of HDAC activity was detected in PC-3 cells. In contrast, DIM significantly inhibited HDAC activity in both cell lines by as much as 66%. Decreases in HDAC activity correlated with increased expression of p21, a known target of HDAC inhibitors. DIM treatment caused a significant decrease in the expression of HDAC2 protein in both cancer cell lines but no significant change in the protein levels of HDAC1, HDAC3, HDAC4, HDAC6 or HDAC8 were detected. Taken together, these results show that inhibition of HDAC activity by DIM may contribute to the phytochemicals anti-proliferative effects in the prostate. The ability of DIM to target aberrant epigenetic patterns, in addition to its effects on detoxification of carcinogens, may make it an effective chemopreventive agent by targeting multiple stages of prostate carcinogenesis.

Rare diseases typically affect fewer than 200,000 patients annually, yet because thousands of rare diseases exist, the cumulative impact is millions of patients worldwide. Every form of childhood cancer qualifies as a rare disease-including the childhood muscle cancer, rhabdomyosarcoma (RMS). The next few years promise to be an exceptionally good era of opportunity for public-private collaboration for rare and childhood cancers. Not only do certain governmental regulation advantages exist, but these advantages are being made permanent with special incentives for pediatric orphan drug-product development. Coupled with a growing understanding of sarcoma tumor biology, synergy with pharmaceutical muscle disease drug-development programs, and emerging publically available preclinical and clinical tools, the outlook for academic-community-industry partnerships in RMS drug development looks promising.

Epidemiological studies provide evidence that consumption of cruciferous vegetables, like broccoli, can reduce the risk of cancer development. Sulforaphane (SFN) is a phytochemical derived from cruciferous vegetables that induces anti-proliferative and pro-apoptotic responses in prostate cancer cells, but not in normal prostate cells. The mechanisms responsible for this cancer-specific cytotoxicity remain unclear. To examine this issue we utilized RNA sequencing and determined the transcriptomes of normal prostate epithelial cells, androgen-dependent prostate cancer cells, and androgen-independent prostate cancer cells treated with SFN. SFN treatment dynamically altered gene expression and resulted in distinct transcriptome profiles depending on prostate cell line. SFN also down-regulated the expression of genes that were up-regulated in prostate cancer cells. Network analysis of genes altered by SFN treatment revealed that the transcription factor Specificity protein 1 (Sp1) was present in an average of 90.5% of networks. Sp1 protein was significantly decreased by SFN treatment in prostate cancer cells and Sp1may be an important mediator of SFN-induced changes in expression. Overall, the data show that SFN alters gene expression differentially in normal and cancer cells with key targets in chemopreventive processes, making it a promising dietary anti-cancer agent.

The dysregulation of long intergenic non‐coding RNA’s (lincRNAs) has become an emerging factor in cancer development. Dietary chemopreventive compounds, such as sulforaphane (SFN) from cruciferous vegetables, can alter epigenetic targets for cancer prevention but the impact of diet on lincRNA expression is unknown. Normal prostate epithelial cells and prostate cancer cells were treated with 15 μM SFN to test the hypothesis that SFN alters the expression of lincRNAs. RNA‐sequencing revealed that hundreds of lincRNAs were differentially expressed in prostate cancer cells relative to normal prostate epithelial cells. SFN treatment significantly altered the expression of ~70 lincRNAs in each cell type. The SFN‐induced response differed depending on cell line and was dynamic over time. In prostate cancer cells, SFN treatment reversed the aberrant expression of some lincRNAs. Preliminary guilt by association work showed that lincRNA's altered by SFN treatment correlate with genes that regulate angiogenesis, cell proliferation, migration, differentiation, apoptosis, and the glutathione metabolic process. Studies evaluating the functional role of these lincRNAs in prostate cancer development are ongoing. Ultimately, this data revealed a novel mechanism by which SFN impacts prostate cancer that may be leveraged to develop future successful prostate cancer prevention strategies. Funding: P01CA090890 Grant Funding Source: Supported by P01CA090890

Sulforaphane (SFN), a phytochemical derived from cruciferous vegetables, induces anti‐proliferative and pro‐apoptotic responses in prostate cancer cells, but not normal prostate cells. The mechanisms responsible for these specific chemopreventive properties remain unclear. We utilized RNA sequencing (RNA‐seq) to test the hypothesis that SFN modifies patterns of gene expression that are critical in prostate cancer progression. Normal prostate epithelial cells and prostate cancer cells were treated with 15 uM SFN and the transcriptome was determined at 6 and 24h time points. SFN alters the mRNA expression levels of ~3,000 genes in each sample and the transcriptional response is highly dynamic over time. SFN treatment also alters the expression of non‐coding RNAs including lincRNAs. SFN targets multiple pathways in normal and prostate cancer cells that are critical in cancer including proliferation, apoptosis, angiogenesis, DNA damage response, and cell migration. Network analysis suggests that the transcription factor Sp1 may be a major mediator by which SFN treatment induces changes in gene expression. Studies evaluating the effect of SFN on Sp1 are ongoing and Sp1 mRNA levels are significantly decreased by SFN treatment. Together, this information can be leveraged to help develop intervention strategies to significantly reduce the incidence and severity of prostate cancer. Funding: P01CA090890Grant Funding Source: National Institute of Health