Mol Cell Oncol. ;2(4):e (-)-Oleocanthal rapidly and selectively induces cancer cell death via lysosomal membrane permeabilization. LeGendre . We investigated the effect of OC on human cancer cell lines in culture and found that OC induced cell death in all cancer cells examined as rapidly as J Cancer. Jul 5;8(11) doi: /jca eCollection of CTCF in Breast Cancer Cells Selectively Induces Cancer Cell Death via p
Cell Induces Death Cancer
All results excluded the induction of parossistic autophagy by GEV. Rather, the robust autophagic flux of A cells was abrogated by this treatment Figure 4B and Supplementary Figure 2.
Effect of glucoevatromonoside on autophagic flux. A Representative transmission electron microscopy of A cells treated with GEV 50 nM with and without bafilomycin A1 10 nM after 12 and 24 h of treatment. The blots shown are representative of three independent experiments. Results were confirmed by real-time imaging Supplementary Figure 3 and Supplementary Videos 6— This result confirms caspase-dependency involved in PS exposure and points at two possible alternative scenarios where apoptotic and non-apoptotic features of cell death might co-exist in different cell subpopulations or the same cell.
Figure 5A shows a reduction of the proliferation of U cells in a concentration-dependent manner, similarly to A cells. GEV-treated U cells undergo changes in their nuclear morphology in a concentration-dependent manner leading to apoptotic nuclear condensation and fragmentation Figures 5C,D.
Altogether, these results indicate the ability of GEV of triggering a caspase-dependent apoptosis in U cells. Glucoevatromonoside impairs cell growth and induces caspase-dependent apoptosis in U A GEV reduces proliferation of U cells in a concentration-dependent manner; B Analysis of cell death performed by trypan blue staining after 24 and 48 h of treatment at 10, 50, and nM; C GEV induces apoptotic nuclear morphology.
Arrows show fragmented nuclei; D Quantification of cell death by Hoechst staining of fragmented nuclei in treated cells vs. It also down-regulated p53 at all concentrations tested as earlier as 12 h Figure 6.
Glucoevatromonoside induces cell cycle arrest. Cell cycle analysis of cells treated with GEV for 24 and 48 h. DMSO served as vehicle control. We then confirmed the potential of GEV to impair the ability of A cells in in vitro and in vivo 3D tumor formation assays in the presence of increasing concentrations of the compound.
To extend our colony formation assays to an in vivo situation, we assessed the capacity of GEV to abrogate A tumor formation in a zebrafish xenograft model. Our results confirmed a strong abrogation of tumor formation after injection of 50 nM GEV-pretreated, fluorescently stained A, validating our in vitro results Figure 7C. Glucoevatromonoside abrogates tumor formation in 3D culture and zebrafish xenografts.
Quantification of tumor area and volume are shown; B A colony formation assays in the presence of increasing concentrations of GEV 10 days. CM-dil-stained A cells were treated for 30 h with 0, 10, 50, and nM GEV and injected in the zebrafish yolk sac at 48 hpf. Representative images from at least 10 fish per condition. PBS injection served as a negative control.
Fluorescent tumor mass was quantified and reported in the graph. The potential of CGs as anticancer drugs arose from the epidemiological evidence of a lower mortality rate of patients suffering from both congestive heart failure and breast cancer under treatment with digoxin Haux, ; Stenkvist, Since then, anticancer activity of CGs have been evaluated in vitro Newman et al.
Repositioning of this class of clinically-validated drugs could allow reducing cancer drug development time and costs Duenas-Gonzalez et al. To validate the putative pharmacological target of GEV, we implemented docking analyses using the Autodock Vina program Trott and Olson, The predicted affinity energies of GEV bound to ouabain and bufalin complexed structures were similar to those of their original ligands, but predicted binding energy of digoxin was higher than that of GEV.
GEV was able to trigger caspase-dependent and -independent cell death mechanisms, depending on the cell context. Of note, GEV presented an interesting differential toxicity. Furthermore, in vitro 3D cell culture using clonogenic and spheroid formation assays as well as in vivo investigations on zebrafish xenograft validated our findings, providing a pre-clinical favorable profile for GEV as new anti-cancer agent.
Overall, lung cancer cell models with different genetic backgrounds were very sensitive, encouraging further investigations on this type of cancer. Our results are in line with previous investigations showing the ability for a same CG to induce cytotoxic effects via execution of different forms of cell death, including UNBS, oleandrin, and digoxin Mijatovic et al. UNBS, for example, could induce autophagy in lung, prostate, pancreatic, and brain cancer cell lines Mijatovic et al.
Interesting prototypical inhibitors of controlled necrosis were unable to protect cells against GEV excluding these alternative mechanisms of cell death induction. Surprisingly, GEV prompts the exposure of phosphatidylserine on the outer leaflet of the plasma membrane in A cells correlated to caspase involvement, shown by zVAD-sensitive PS exposure. CGs displayed anti-proliferative activity via cell cycle inhibition Mijatovic et al.
Our results are in line with Wang and coworkers Wang et al. Consequently, cyclin B1 expression was increased after GEV treatment at 10 nM followed by a decrease at highest concentrations.
Elevated cyclin B1 expression levels are a common feature of various cancer models Dutta et al. Cyclin B-cyclin dependent kinase CDK complexes are active regulators of the progression of the cell cycle as they initiate and maintain the phosphorylated state of the retinoblastoma protein pRb , which plays a central role in proliferation. We also observed that p53 was downregulated at all concentrations tested.
This tumor suppressor controls multiple cell cycle checkpoints regulating the mammalian response to DNA damage. Nakayama and Yamaguchi Nakayama and Yamaguchi, suggested that the reduction of cyclin B1 levels may lead to the restoration of the p53 pathway, but not when A cells were treated with GEV. Also, cells died independently of cyclin B1 and p53 status. These results agree with the findings of Wang et al. Moreover, these effects occurred preferentially in cancerous cells vs.
In conclusion, GEV reduced the proliferation and the viability of various cell types. Nevertheless, GEV triggered a non-canonical mode of cell death as caspase-mediated PS exposure was identified in selected sub-populations. In a leukemia model, GEV led to caspase-dependent apoptosis concomitant with an accumulation of cells in subG0. Overall, our results show the cytotoxic effects of GEV on A and U cells by inducing non-canonical cell death in solid tumor cells and canonical cell death in human leukemia.
The solid tumor A cells exhibit a very high basal autophagic flux, compatible with their high proliferation rate. GEV does not further exacerbate autophagy, rather this compound seems may have some potentials to impair it, for example looking at the reduced accumulation of LC3I-II at earlier times of treatment and lover concentrations, a condition which was not prevented by bafilomycin A1.
Inhibition of autophagy exacerbates cell death induction as a critical mechanism for cellular energy and nutrition homeostasis, as well as cell survival in many forms of cancer. As such, it could be interesting in the future to verify if GEV might be especially promising against autophagy-addicted cancer types characterized by improved stress and chemotherapy resistance.
This study was carried out in accordance and with the agreement of the recommendations of the SNU College of Pharmacy animal welfare legislation. Participated in research design: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The Supplementary Material for this article can be found online at: Antiherpes activity of glucoevatromonoside, a cardenolide isolated from a Brazilian cultivar of Digitalis lanata.
Isolation of cardenolides from a Brazilian cultivar of Digitalis lanata by rotation locular counter-current chromatography. Subapoptogenic oxidative stress strongly increases the activity of the glycolytic key enzyme glyceraldehyde 3-phosphate dehydrogenase. Assembling the puzzle of anti-cancer mechanisms triggered by cardiac glycosides.
New horizons for old drugs and drug leads. Synthetic polysulfane derivatives induce cell cycle arrest and apoptotic cell death in human hematopoietic cancer cells. Non-canonical programmed cell death mechanisms triggered by natural compounds. Valproic acid as epigenetic cancer drug: Cyclins as markers of tumor proliferation: Digitoxin and a synthetic monosaccharide analog inhibit cell viability in lung cancer cells.
Features and development of Coot. Silver nanoparticles synthesized from Adenium obesum leaf extract induced DNA damage, apoptosis and autophagy via generation of reactive oxygen species. B Biointerfaces , — Prognostic value of cyclin B1 protein expression in colorectal cancer. Digitoxin is a potential anticancer agent for several types of cancer.
First-in-human study of pbi, an oleander-derived inhibitor of akt, fgf-2, nf-kappaBeta and p70s6k, in patients with advanced solid tumors. New Drugs 32, — Targeting FXYD2 by cardiac glycosides potently blocks tumor growth in ovarian clear cell carcinoma. Digoxigenin modification of adenovirus to spatially control gene delivery from chitosan surfaces. Concurrent detection of autolysosome formation and lysosomal degradation by flow cytometry in a high-content screen for inducers of autophagy.
UNBS, a steroid cardiac glycoside inducing apoptotic cell death in human leukemia cells. Lanatoside C suppressed colorectal cancer cell growth by inducing mitochondrial dysfunction and increased radiation sensitivity by impairing DNA damage repair.
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Phase 1 trial of Anvirzel in patients with refractory solid tumors. New Drugs 24, — Cardiac glycosides exert anticancer effects by inducing immunogenic cell death. The cardenolide UNBS is able to deactivate nuclear factor kappaB-mediated cytoprotective effects in human non-small cell lung cancer cells. Natural products as sources of new drugs from to Autophagic cell death of human pancreatic tumor cells mediated by oleandrin, a lipid-soluble cardiac glycoside.
Cardiac glycosides as novel cancer therapeutic agents. Monosaccharide digitoxin derivative sensitize human non-small cell lung cancer cells to anoikis through Mcl-1 proteasomal degradation.
Digitoxin-induced cytotoxicity in cancer cells is mediated through distinct kinase and interferon signaling networks. Cell type-dependent ROS and mitophagy response leads to apoptosis or necroptosis in neuroblastoma. Sustained exposure to the DNA demethylating agent, 2'-deoxyazacytidine, leads to apoptotic cell death in chronic myeloid leukemia by promoting differentiation, senescence, and autophagy. Natural compounds as inflammation inhibitors.
Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal omega. Cardiac glycosides in cancer therapy: Such suicide — or apoptosis — occurs by activating a specific program within the cells, leading to their destruction without damaging surrounding cells and tissues.
Apoptosis is crucial during embryonic development and in growth of adult tissues, e. Menstruation — sloughing of the inner lining of the uterus — requires apoptosis. Our body routinely entrusts apoptosis to eliminate faulty white blood cells that could cause autoimmune diseases, or to eliminate abnormal cells which could lead to cancer or leukemias.
The development of abnormal cells into cancer is dependent on several factors, one of which is their ability to escape apoptosis. Cancer cells not only multiply uncontrollably, they successfully dodge this natural mechanism of death. Cancer cells hijack the cell machinery and stop apoptosis by blocking the activity of a particular gene — p This protector gene first stops the division of abnormal cells, and then initiates either their repair or destruction if the damage is severe.
Normally, the p53 gene activates specific apoptotic proteins from the Bcl2 family and the Caspases enzymes that initiate and continue the process of apoptosis. The malfunctioning of the p53 gene results in rapidly growing and aggressive cancers. Restoration of the normal process of apoptosis is one of the avenues being explored for anticancer treatments.
Several drugs, including Aspirin and other non-steroidal-anti-inflammatory-drugs NSAIDs , are being researched in cancer cells for their usage as apoptosis inducers. We explored the potential of micronutrients in inducing apoptosis in different cancer cell lines. Cancer cells are typically immortal, but our results show that a specific micronutrient combination is capable of triggering metabolic and genetic changes that kill cancer cells by inducing their natural cell death cycle.
Interestingly, GEV triggered caspase-dependent apoptosis in U acute myeloid leukemia cells, witnessing cancer-type specific cell death induction. In this study, we demonstrate that a combination of quinacrine and SAHA is potent in inducing cancer cell death and abrogating tumor growth. Amicoumacin A is an antibiotic that was recently shown to target bacterial ribosomes. It affects translocation and provides an additional contact.