Interaction of Human Tumor Cells with Human Platelets and the Coagulation System

SummaryThis paper reports studies on the interaction between human platelets, the plasma coagulation system, and two human tumor cell lines grown in tissue culture: Melanoma and breast adenocarcinoma. The interaction was monitored through the use of 125I- labelled fibrinogen, which measures both thrombin activity generated by cell-plasma interaction and fibrin/fibrinogen binding to platelets and tumor cells. Each tumor cell line activates both the platelets and the coagulation system simultaneously resulting in the generation of thrombin or thrombin-like activity. The melanoma cells activate the coagulation system through “the extrinsic pathway” with a tissue factor-like effect on factor VII, but the breast tumor seems to activate factor X directly. Both tumor cell lines activate platelets to “make available” a platelet- derived procoagulant material necessary for the conversion of prothrombin to thrombin. The tumor-derived procoagulant activity and the platelet aggregating potential of cells do not seem to be inter-related, and they are not specific to malignant cells.

Download Full-text

Abstract 1456:In vitroproduction of circulating tumor cells (CTCs) using 3D cultures of human tumor tissues and established tumor cell lines.

10.1158/1538-7445.am2013-1456 ◽

2013 ◽

Author(s):

William P. Pfund ◽

George Martin ◽

Paul A. Neeb

Keyword(s):

Tumor Cell ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Cell Lines ◽

Human Tumor ◽

Tumor Cell Lines ◽

3D Cultures ◽

Established Tumor ◽

Tumor Tissues

Download Full-text

Unraveling the functional relevance of the Ubiquitin-specific peptidase 22 for necroptotic cell death

10.21248/gups.60925 ◽

2021 ◽

Author(s):

◽

Jens Rödig

Keyword(s):

Cell Death ◽

Programmed Cell Death ◽

Tumor Cell ◽

Tumor Cells ◽

Cell Lines ◽

Human Tumor ◽

Tumor Cell Lines ◽

Human Tumor Cell ◽

Human Tumor Cell Lines ◽

Cell Death Signaling

Ubiquitination is regarded as one of the key post-translational modifications in nearly all biological processes, endowed with numerous layers of complexity. Deubiquitinating enzymes (DUBs) dynamically counterbalance ubiquitination events by deconjugating ubiquitin signals from substrates. Dysregulation of the ubiquitin code and its negative regulators drive various pathologies, such as neurological disorders and cancer. The DUB ubiquitin-specific peptidase 22 (USP22) is well-known for its essential role in the human Spt-Ada-Gcn5 acetyltransferase (SAGA) complex, mediating the removal of monoubiquitination events from Histone 2A and 2B (H2A and -B), thereby regulating gene transcription. In cancer, USP22 was initially described as a part of an 11-gene expression signature profile, predicting tumor metastasis, reoccurrence and death after therapy in a wide range of tumor cells. However, novel roles for USP22 have emerged recently, accrediting USP22 essential roles in regulating tumor development as well as apoptotic cell death signaling. One of the hallmarks of cancer is the evasion of cell death, especially apoptosis, a form of programmed cell death (PCD). Necroptosis, a regulated form of necrosis, is regarded as an attractive therapeutic strategy to overcome apoptosis-resistance in tumor cells, although a profound understanding of the exact signaling cascade still remains elusive. Nevertheless, several ubiquitination and deubiquitination events are described in fine-tuning necroptotic signaling. In this study, we describe a novel role for USP22 in regulating necroptotic cell death signaling in human tumor cell lines. USP22 depletion significantly delayed TNFa/Smac mimetic/zVAD.fmk (TBZ)-induced necroptosis, without affecting TNFa-induced nuclear factor-kappa B (NF-KB) signaling or TNFa-mediated extrinsic apoptosis. Intriguingly, re-expression of USP22 wildtype in the USP22 knockout background could re-sensitize HT-29 cells to TBZ-induced necroptosis, whereas re-constitution with the catalytic inactive mutant USP22 Cys185Ser did not rescue susceptibility to TBZ-induced necroptosis, confirming the USP22 DUB-function a pivotal role in regulating necroptotic cell death. USP22 depletion facilitated ubiquitination and unexpectedly also phosphorylation of Receptor-interacting protein kinase 3 (RIPK3) during necroptosis induction, as shown by Tandem Ubiquitin Binding Entities (TUBE) pulldowns and in vivo (de)ubiquitination immunoprecipitations. To substantiate our findings, we performed mass-spectrometric ubiquitin remnant profiling and identified the three novel USP22-regulated RIPK3 ubiquitination sites Lysine (K) 42, K351 and K518 upon TBZ-induced necroptosis. Further assessment of these ubiquitination sites unraveled, that mutation of K518 in RIPK3 reduced necroptosis-associated RIPK3 ubiquitination and additionally affected RIPK3 phosphorylation upon necroptosis induction. At the same time, genetic knock-in of RIPK3 K518R sensitizes tumor cells to TNFa-induced necroptotic cell death and amplified necrosome formation. In summary we identified USP22 as a new regulator of TBZ-induced necroptosis in various human tumor cell lines and further unraveled the distinctive role of DUBs and (de)ubiquitination events in controlling programmed cell death signaling.

Download Full-text

Synthesis, Spectroscopic Characterization, Structural Studies, and In Vitro Antitumor Activities of Pyridine-3-carbaldehyde Thiosemicarbazone Derivatives

Journal of Chemistry ◽

10.1155/2020/2960165 ◽

2020 ◽

Vol 2020 ◽

pp. 1-12

Author(s):

Wilfredo Hernández ◽

Fernando Carrasco ◽

Abraham Vaisberg ◽

Evgenia Spodine ◽

Jorge Manzur ◽

...

Keyword(s):

Tumor Cell ◽

Cell Lines ◽

Human Tumor ◽

Breast Adenocarcinoma ◽

Significant Activity ◽

Tumor Cell Lines ◽

Human Tumor Cell ◽

Human Tumor Cell Lines ◽

Biological Studies

Eight new thiosemicarbazone derivatives, 6-(1-trifluoroethoxy)pyridine-3-carbaldehyde thiosemicarbazone (1), 6-(4′-fluorophenyl)pyridine-3-carbaldehyde thiosemicarbazone (2), 5-chloro-pyridine-3-carbaldehyde thiosemicarbazone (3), 2-chloro-5-bromo-pyridine-3-carbaldehyde thiosemicarbazone (4), 6-(3′,4′-dimethoxyphenyl)pyridine-3-carbaldehyde thiosemicarbazone (5), 2-chloro-5-fluor-pyridine-3-carbaldehyde thiosemicarbazone, (6), 5-iodo-pyridine-3-carbaldehyde thiosemicarbazone (7), and 6-(3′,5′-dichlorophenyl)pyridine-3-carbaldehyde thiosemicarbazone (8) were synthesized, from the reaction of the corresponding pyridine-3-carbaldehyde with thiosemicarbazide. The synthesized compounds were characterized by ESI-Mass, UV-Vis, IR, and NMR (1H, 13C, 19F) spectroscopic techniques. Molar mass values and spectroscopic data are consistent with the proposed structural formulas. The molecular structure of 7 has been also confirmed by single crystal X-ray diffraction. In the solid state 7 exists in the E conformation about the N2-N3 bond; 7 also presents the E conformation in solution, as evidenced by 1H NMR spectroscopy. The in vitro antitumor activity of the synthesized compounds was studied on six human tumor cell lines: H460 (lung large cell carcinoma), HuTu80 (duodenum adenocarcinoma), DU145 (prostate carcinoma), MCF-7 (breast adenocarcinoma), M-14 (amelanotic melanoma), and HT-29 (colon adenocarcinoma). Furthermore, toxicity studies in 3T3 normal cells were carried out for the prepared compounds. The results were expressed as IC50 and the selectivity index (SI) was calculated. Biological studies revealed that 1 (IC50 = 3.36 to 21.35 μM) displayed the highest antiproliferative activity, as compared to the other tested thiosemicarbazones (IC50 = 40.00 to >582.26 μM) against different types of human tumor cell lines. 1 was found to be about twice as cytotoxic (SI = 1.82) than 5-fluorouracile (5-FU) against the M14 cell line, indicating its efficiency in inhibiting the cell growth even at low concentrations. A slightly less efficient activity was shown by 1 towards the HuTu80 and MCF7 tumor cell lines, as compared to that of 5-FU. Therefore, 1 can be considered as a promising candidate to be used as a pharmacological agent, since it presents significant activity and was found to be more innocuous than the 5-FU anticancer drug against the 3T3 mouse embryo fibroblast cells.

Download Full-text

Cytokine profile of conditioned medium from human tumor cell lines after acute and fractionated doses of gamma radiation and its effect on survival of bystander tumor cells

Cytokine ◽

10.1016/j.cyto.2012.08.022 ◽

2013 ◽

Vol 61 (1) ◽

pp. 54-62 ◽

Cited By ~ 71

Author(s):

Sejal Desai ◽

Amit Kumar ◽

S. Laskar ◽

B.N. Pandey

Keyword(s):

Gamma Radiation ◽

Tumor Cell ◽

Conditioned Medium ◽

Tumor Cells ◽

Cell Lines ◽

Human Tumor ◽

Cytokine Profile ◽

Tumor Cell Lines ◽

Human Tumor Cell ◽

Human Tumor Cell Lines

Download Full-text

Phosphodiesterase 10, a novel target in colon cancer.

Journal of Clinical Oncology ◽

10.1200/jco.2014.32.3_suppl.503 ◽

2014 ◽

Vol 32 (3_suppl) ◽

pp. 503-503

Author(s):

Suzanne Russo ◽

Nan Li ◽

Kevin Lee ◽

Yaguang Xi ◽

Bing Zhu ◽

...

Keyword(s):

Colon Cancer ◽

Mouse Model ◽

Tumor Cell ◽

Tumor Cells ◽

Cell Lines ◽

Human Tumor ◽

Clinical Samples ◽

Mrna Levels ◽

Tumor Cell Lines ◽

Sirna Knockdown

503 Background: Elevation of intracellular cGMP is known to inhibit tumor proliferation and induce apoptosis, although the phosphodiesterase (PDE) isozymes that regulate cGMP levels in tumor cells have not been well studied. We report first evidence that PDE10 is elevated in colon tumors compared with normal colon and suggest that PDE10 inhibitors can be used for the treatment or prevention of colon cancer. Methods: PDE10 protein and mRNA levels were measured in human colon tumor cells (HT29, HCT116, SW480, Caco2), normal colonocytes (NCM460), human clinical samples, and ApcMin/+ mouse model. Two chemically distinct PDE10 selective inhibitors, PQ-10 and Pf-2545920, were tested against the cell lines. The NCI-60 panel of human tumor cell lines was also screened against Pf-2545920 to identify potential differences in sensitivity among histologically diverse tumor types. We also performed siRNA knockdown studies in colonocytes and tumor cell lines. To determine the effect of the PDE10 siRNA knockdown on cyclic nucleotide hydrolysis, whole cell lysates from transfected cells were assayed for PDE activity using cGMP or cAMP as substrates. Results: PDE10 levels were low in normal colonocytes (NCM460) and elevated in tumor cell lines. Similarly, PDE10 was elevated human clinical specimens and the ApcMin+/ mouse model compared with normal mucosa. PDE10 inhibitors and siRNA selectively inhibited colonic tumor growth while stable knockdown inhibited colony formation and increased doubling time. Pf-2545920 also supressed growth of all cell lines within the NCI-60 panel. In comparison with lysates from vector control cells, transfection with PDE10 siRNA reduced cGMP hydrolysis by ~35% in both HCT116 and HT29 cell lines, but did not affect cGMP hydrolysis in colonocytes; siRNA did not significantly affect cAMP degradation in all 3 cell lines. Conclusions: PDE10 plays a role in colon tumorgenesis whereby inhibitors can selectively suppress tumor cell growth. The mechanism by which PDE10 inhibition affects growth appears to involve activation of cGMP/PKG signalling. PDE10 represents a novel anticancer target for the treament and prevention of colon cancer.

Download Full-text

Expression of rat complement control protein Crry on tumor cells inhibits rat natural killer cell–mediated cytotoxicity

Blood ◽

10.1182/blood.v100.9.3304 ◽

2002 ◽

Vol 100 (9) ◽

pp. 3304-3310 ◽

Cited By ~ 15

Author(s):

Theresa A. Caragine ◽

Masaki Imai ◽

Alan B. Frey ◽

Stephen Tomlinson

Keyword(s):

Tumor Cell ◽

Nk Cells ◽

Natural Killer ◽

Tumor Cells ◽

Cell Lines ◽

Nk Cell ◽

Human Tumor ◽

Mammary Adenocarcinoma ◽

Tumor Cell Lines ◽

Human Tumor Cell

Abstract Crry is a rodent membrane–bound inhibitor of complement activation and is a structural and functional analog of the human complement inhibitors decay-accelerating factor and membrane cofactor protein. We found previously that expression of rat Crry on a human tumor cell line enhances tumorigenicity in nude rats. In this study, we investigated the effect that rat Crry expressed on tumor cells has on rat cell–mediated cytotoxicity and antibody-dependent cell-mediated cytotoxicity (ADCC). The expression of rat Crry on the surface of different human tumor cell lines inhibited ADCC mediated by rat natural killer (NK) cells. C3 opsonization is known to enhance NK cell–mediated cytolysis, and a potential mechanism for Crry-mediated inhibition of NK cell lysis is through Crry modulation of C3 deposition on target cells. However, the transfection of tumor cell lines with Crry enhanced their resistance to NK cell–mediated lysis in the absence of exogenous complement. The resistance of Crry-expressing tumor cells to NK cell–mediated ADCC could be reversed by treatment with anti–Crry F(ab)2. In addition, anti–Crry F(ab)2 enhanced the susceptibility of 13762 rat mammary adenocarcinoma cells (that endogenously express Crry) to ADCC mediated by allogeneic rat NK cells in the absence of added complement. We found no evidence that rat NK cells were a source of complement for target cell deposition during the in vitro cytolysis assay. These data suggest a novel function for rat Crry in tumor immune surveillance that may be unrelated to complement inhibition.

Download Full-text

Synthesis and antiproliferative activity of various novel indole Mannich bases

Advances in Modern Oncology Research ◽

10.18282/amor.v1.i2.4 ◽

2015 ◽

Vol 1 (2) ◽

Cited By ~ 2

Author(s):

Mardia T El Sayed ◽

Hoda A R Hussein ◽

Khadiga M Ahmed ◽

Nehal A Hamdy

Keyword(s):

Tumor Cell ◽

Cell Lines ◽

Cancer Cell ◽

Antiproliferative Activity ◽

Human Tumor ◽

Inhibitory Effect ◽

Mannich Bases ◽

Primary Amines ◽

Breast Adenocarcinoma ◽

Tumor Cell Lines

Various secondary and primary amines were converted into bis-indolyl Mannich bases with good to excellent yields via double condensation reactions with indole and glutaraldehyde. The expected bis-indolyl Mannich bases (2, 3 and 4) were formed by using piperazinehexahydrate and 4,4′-trimethylenedipiperidine. Whereas, the use of primary amines, phenylhydrazine, amino acids and primary diamine produced the corresponding bis-indolyl-1,2,6-trisubstituted pipridines (5a-e) and indolyl-quinolizine (6) and dibis-indolyl-1,2,6-trisubstituted pipridines (7). All analytical and spectral data of these bis-indolyl Mannich bases have been determined. Six of the synthesized bis-indolyl Mannich bases have been subjected for antiproliferative activity screening at National Cancer Institute (NCI), Egypt, towards three human tumor cell lines representing different tumor types: breast adenocarcinoma cell (MCF-7), non-small lung cancer cell (NCI-H460), and central nervous system (CNS) cancer cell (SF-268). Compound (4) indicated the best and highest inhibitory effect against all three tested tumor cell lines with inhibition of 50% concentration (IC50) for MCF-7 (0.08 µmol/L), NCI-H460 (0.05 µmol/L) and SF-268 (0.01 µmol/L).

Download Full-text