Epigenetic Targeting of Heparan Sulfate 3- O - and 6- O -Sulfation in Breast Cancer Cells: Prospects for Attenuating Prothrombotic Tumor Cell Activities.
Nico BückreißMarie Schulz-FinckePhilipp KönigMarco MaccaranaToin H van KuppeveltJin-Ping LiMartin GötteGerd BendasPublished in: ACS pharmacology & translational science (2024)
The deregulation of cell surface heparan sulfate proteoglycans (HSPGs) is a main issue of cancer cells for increasing their malignancy. In these terms, the sulfation pattern of HS, created by an orchestrated activity of enzymes balancing a site-specific sulfation, is of key importance. These enzymes are often deregulated by epigenetic processes in cancer, e.g., being silenced by DNA hypermethylation. Here, we address this issue in human breast cancer cell lines aiming to target epigenetic processes to reactivate HS sulfation, shifting HS into an antithrombotic phenotype for which 3- O -sulfation is particularly important. Treatment of MCF-7 and MDA-MB-231 cells with nontoxic concentrations of 5-azacytidine (azacytidine) and 5-fluoro-2'-deoxycytidine (FdCyd) as DNMT inhibitors or vorinostat for targeting HDAC increased HS3- O -sulfation remarkably, as confirmed by fluorescence microscopy, by upregulating HS3- O -sulfotransferases, detected by quantitative real-time polymerase chain reaction and Western blot. Flow cytometry and microscopic approaches confirm that upon inhibitor treatment, increased HS3- O -sulfation improves cell binding to antithrombin, leading to an antithrombotic activity. Nevertheless, only azacytidine- and vorinostat-treated cells display anticoagulative properties, represented by attenuated thrombin formation, a lower activation of human platelet aggregation, or ATP release. In contrast, FdCyd additionally upregulated tissue factor expression in both cell lines, overshadowing the anticoagulant effects of HS, leading to an overall prothrombotic phenotype. Our data provide evidence for the first time that targeting epigenetic processes in HS sulfation is a valuable means to foster anticoagulative cell properties for decreasing malignancy and metastatic potency. These data warrant further investigations to fine-tune epigenetic targeting and to search for potential biomarkers attributed to these activities.
Keyphrases
- dna methylation
- breast cancer cells
- gene expression
- single cell
- induced apoptosis
- cell cycle arrest
- endothelial cells
- cell therapy
- cancer therapy
- single molecule
- flow cytometry
- atrial fibrillation
- squamous cell carcinoma
- small cell lung cancer
- cell surface
- electronic health record
- high resolution
- stem cells
- magnetic resonance imaging
- mesenchymal stem cells
- histone deacetylase
- cell death
- big data
- mass spectrometry
- high throughput
- young adults
- pluripotent stem cells
- machine learning
- deep learning
- air pollution
- bone marrow
- drug delivery
- binding protein
- cell free
- circulating tumor cells
- papillary thyroid