Stilbenoids remodel the DNA methylation patterns in breast cancer cells and inhibit oncogenic NOTCH signaling through epigenetic regulation of MAML2 transcriptional activity

DNA hypomethylation was previously implicated in cancer progression and metastasis. The purpose of this study was to examine whether stilbenoids, resveratrol and pterostilbene thought to exert anticancer effects, target genes with oncogenic function for de novo methylation and silencing, leading to inactivation of related signaling pathways. Following Illumina 450K, genome-wide DNA methylation analysis reveals that stilbenoids alter DNA methylation patterns in breast cancer cells. On average, 75% of differentially methylated genes have increased methylation, and these genes are enriched for oncogenic functions, including NOTCH signaling pathway. MAML2, a coactivator of NOTCH targets, is methylated at the enhancer region and transcriptionally silenced in response to stilbenoids, possibly explaining the downregulation of NOTCH target genes. The increased DNA methylation at MAML2 enhancer coincides with increased occupancy of repressive histone marks and decrease in activating marks. This condensed chromatin structure is associated with binding of DNMT3B and decreased occupancy of OCT1 transcription factor at MAML2 enhancer, suggesting a role of DNMT3B in increasing methylation of MAML2 after stilbenoid treatment. Our results deliver a novel insight into epigenetic regulation of oncogenic signals in cancer and provide support for epigenetic-targeting strategies as an effective anticancer approach

Fertility Counseling Pattern over Time in Young Patients with Breast Cancer: A Retrospective Analysis at a Large Comprehensive Cancer Center

Background: One main issue to be considered in young patients diagnosed with early breast cancer (BC) is the impact of oncological treatments on fertility and future chances of conception. Current guidelines recommend a comprehensive addressing of oncofertility as part of the management of premenopausal BC patients, including counselling on available assisted reproduction technologies and fertility preservation (FP) strategies. The COVID-19 pandemic represented a potential hurdle to the integration of these procedures into clinical practice. This study aims to describe the time-related evolution in addressing oncofertility issues. Methods: This retrospective mono-institutional observational study considered 206 patients who received neoadjuvant chemotherapy, adjuvant chemotherapy (CT) or adjuvant endocrine therapy (ET), diagnosed with breast cancer at the age of 40 or younger in the years 2014-2015 and 2020-2021. Timerelated evolution in addressing oncofertility during oncological consultations and adoption of a fertility or ovarian function preservation (OFP) method were analyzed comparing the two different timeframes. Results: Comparing the two cohorts 2014-2015 and 2020-2021, we found a significant difference in the presence of fertility discussion records (37.4% vs 57.9%, p < 0.01), and in the application of OFP/FP techniques (54.5 vs 78.5%, p < 0.01). In the two cohorts there was a significant difference in OFP (57.6% vs 70%, p = 0.03) and FP techniques application rates (5.1% vs 19.6%, p < 0.01). In the study population, age at diagnosis resulted to influence clinicians' approach towards counseling and/or OFP/FP strategies (87.3% in patients <35 years old (yo) vs 56.7% in older patients, p < 0.01). In the 2020-2021 cohort, age resulted less influential in the choice of using an OFP/FP strategy (87% vs 72.1%, p = 0.18). A higher rate of documented fertility discussion and/or OFP/FP techniques application was recorder in patients who had not had children before BC diagnosis (80.6% vs 64.5%, p = 0.02). When considering only the 2020-2021 timeframe, parity no longer significantly affected the prescription of an OFP/FP strategy (80.4% vs 78.3%, p = 0.93). Conclusions: This study on real world data demonstrates the progressive evolution in the way clinicians approach oncofertility issues, showing a greater attention across years, with more BC patients receiving a dedicated counseling, despite the COVID-19 pandemic

Analysis of neuropeptide gene expression by transfection of DNA into cell lines

The transcriptional regulation of neuropeptide genes by cAMP is often directed by a cAMP responsive enhancer (CRE) upstream to the promoter of the genes. The identity of the CRE was determined by transient transfection experiments and has the consensus sequence of 5′-TGACGTCA-3′. A large family of transcription factors have been identified which recognize the CRE. Transient transfection assays that employed expression of these factors driven by viral promoters have determined they can transactivate the neuropeptide gene promoters. Because of the large number of factors that have been identified as being able to recognize the CRE, it has been difficult to determine which factors mediate in vivo the transactivation of a particular CRE. Using a dominant negative mutant of one of these factors, CREB, it has been determined that both CREB as well as other factors which do not interact with CREB may mediate the cAMP response of the somatostatin gene.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/43232/1/11022_2005_Article_BF01667368.pd