The conjugated antimetabolite 5-FdU-ECyd and its cellular and molecular effects on platinum-sensitive vs. -resistant ovarian cancer cellsin vitro

Abstract

BackgroundResistance to platinum-based chemotherapy is a clinical challenge in the treatment of ovarian cancer (OC) and limits survival. Therefore, innovative drugs against platinum-resistance are urgently needed. Our therapeutic concept is based on the conjugation of two chemotherapeutic compounds to a monotherapeutic pro-drug, which is taken up by cancer cells and cleaved into active cytostatic metabolites. We explore the activity of the duplex-prodrug 5-FdU-ECyd, covalently linking 2'-deoxy-5-fluorouridine (5-FdU) and 3'-C-ethynylcytidine (ECyd), on platinum-resistant OC cells.MethodsIn vitro assays and RNA-Sequencing were applied for characterization of 5-FdU-ECyd treated platinum-sensitive A2780 and isogenic platinum-resistant A2780cis and independent platinum-resistant Skov-3-IP OC cells.ResultsNano molar 5-FdU-ECyd concentrations induced a rapid dose-dependent decline of cell viability in platinum-sensitive and -resistant OC cells. The effect of 5-FdU-ECyd was accompanied by the formation of DNA double strand breaks and apoptosis induction, indicated by a strong increase of pro-apoptotic molecular markers. Moreover, 5-FdU-ECyd efficiently decreased migration of platinum-resistant OC cells and inhibited clonogenic or spheroidal growth. Transcriptome analysis showed early up-regulation of CDKN1A and c-Fos in both, platinum-resistant and -sensitive cells after 5-FdU-ECyd treatment and de-regulation of distinct cellular pathways involved in cell cycle regulation, apoptosis, DNA-damage response and RNA-metabolism. Combined treatment of 5-FdU-ECyd and cisplatin did not show a synergistic cellular response, suggesting the potential use of 5-FdU-ECyd as a monotherapeutic agent.ConclusionOur data provide novel mechanistic insight into the anti-tumor effect of 5-FdU-ECyd and we hypothesize that this duplex-prodrug could be a promising therapeutic option for OC patients with resistance to platinum-based chemotherapy.

Functional Analysis of Non-Genetic Resistance to Platinum in Epithelial Ovarian Cancer Reveals a Role for the MBD3-NuRD Complex in Resistance Development

“…2.12. Analysis of RNA-Seq Data of EOC Cell Lines and Patient Samples RNA-seq data were obtained from a previously published dataset for A2780 and A2780cis cells (GSE98230) [31]. Raw counts of A2780 and A2780cis cells were extracted and processed on the galaxy platform (https://usegalaxy.eu/; accessed on 26 July 2021) [25].…”

Hypoxia-induced epigenetic silencing of polo-like kinase 2 promotes fibrosis in atrial fibrillation

“…RNA sequencing and bioinformatics. The RNA sequencing and subsequent bioinformatical analysis was done as published by Schott et al (Schott et al, 2017). For Whole-Transcriptome Sequencing (RNA-Seq), 1.5x 10 6 murine cardiac PLK2 WT and KO fibroblasts were seeded in T-25 flasks and cultured until optical confluency.…”

Time-Dependent Effects of POT1 Knockdown on Proliferation, Tumorigenicity, and HDACi Response of SK-OV3 Ovarian Cancer Cells

“…The treatment of ovarian cancer is difficult for clinicians and researchers who work in the field of oncology. Some targeted therapies are already approved for ovarian cancer among the treatment of primary or recurrent disease, such as antiangiogenic therapy with bevacizumab or PARP inhibitors [ 14 , 15 ]. Analyses of available data suggest that HDACis exert anticancer effects by specifically targeting transcription factors [ 16 ] and promoting deacetylation changes in these nonhistone protein substrates.…”